岡山醫學會雜誌第四百十二號大正十三年五月三十一日發行

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(1)岡山醫學會雜誌第四百十二號大正十三年五月三十一日發行. 原 A Contribution. 著 to the. Bone-regeneration:. with Observations. upon. of Periosteum. the and. Transplantation Bone-marrow.. By Dr. Koken. Ito.. From the Pathological Department of Okayama Medical College. (Director: Prof. Dr. O. Tamura.). Table I. II.. Contents. VI.. Introduction. Literature.. III.. Methode,. I V.. Experiments. V.. of. VII. VIII.. Material. Nos.. one to fifteen.. IX.. Summary.. X.. I.. Appendix. Discussion. Conclusion. Bibliography. Description. of plates.. Introduction.. Our knowledge of bone regeneration began about the eighteenth cen tury and since that time there have been many reports about the experi mental and clinical sides of the subject. Bone transplantation is now be coming an important branch of surgery, however, no definite conclusions as to the real origin of the bone regeneration have been reached. Generally, in the primary ossification in the embryo is divided into two parts: 1. Intramembranous, direct, or metaplastic ossification,-this development comes from the embryonal connective tissue,-the mesenchym..

(2) 2. -424-. 2.. Intracartilaginous,. indirect, or neoplastic ossification,-hyaline. cartilage,. which is also derived from the embryonal connective tissue. In both types, the actual processes of ossification are essentially the same and the bone is completed by the cooperation of the periosteum. is hitherto believed that the bone regeneration. It. is effected by osteoblasts. which give rise to new bone. These osteoblasts in children and adoles cents may apparently be derived either from the marrow or from the perios teum, but in adults cheifly, if not wholly, from the periosteum.. This peri. osteum is a connective tissue membrane covering the external surface of all the bone except the cartilage in the joint and it consists of two layers: an outer layer of dense connective tissue, rich in blood vessels and containing also lymphatic vessels and nerves, and an inner layer which poor in blood vessels but has an abundance of elastic fibers. The cells of the inner layer of the periosteum are spindle shaped or flattened connective tissue cells, to gether with the more cuboidal osteoblasts which rest against the bone.. In. young bone there are so numerous as to form a third layer of the perio steum. In adults they are few in number, but are capable of proliferation, and together with those in the endosteum, they are the source of new bone after destruction of its osseous tissue. It is quite difficult to distinguish microscopically on section, the bounda ry line of the periosteum and the real bone, also the outer layer of the periosteum blends with the surrounding looser connective tissue and in places with fasciae and tendons. Whether the periosteum as well as the endos teum is the main agent in bone regeneration or not, is a matter of discus sion.. I briefly quote some literature as to the subject:-. II. Duhamel1. 1739.. long bone of young covered. He placed animal. Literature. a silver ring. 1858.. the. periosteum. of a. and after a while, he found that it had become. by the bone and so he believed. periosteum. Virchow2. under. He claimed. that the. bone was. derived. that the bone cell was derivered. from. from the.

(3) -425-. periosteum by the process of metaplasia. Gegenbauer3 gave the term "Osteoblast" and he agreed with the above view. Ollier4 1867. His theory is that all transplanted bone with the peri osteum, retains its inherent activity and the bone denuded its periosteum, undergoes necrosis and is completely absorbed. Moschcowitz51895. He pointed out, based upon the study of cal cification and ossification of the ovary that osteoblast is simply an endo thelial cell from the lining of the blood vessel. He also said that ossi fi cation does not occur without preliminary calcificationand calcificationoc curs only in dead tissue. Macewen6 1912. He said that the periosteum is of great use in limi ting, within specific boundaries, the distribution of the osteoblasts, and pre venting them during their evolutionary period from being scattered into the soft tissue, where their presence would be prejudicial to the function of these parts. In the loose areolar tissue existing between the periosteum and the bone, the osteoblasts find nutrient for their growth and also space to generate, free from undue pressure. While not under-estimating the periosteum is a limiting and protection membrane, of great use in physio logical and pathological conditions, there are no data to indicate that it can, of itself, secrete or reproduce bone. He claimed that in bone growth, diaphyseal osteoblasts are generated from the nuclei of the diaphyseal car tilage cells. This diaphyseal cartilage is but a phase in the evolution of the bone cells. When the nuclei of the cartilage cells proliferate, the car tilaginous envelopes become less distinct and finally disappear, the space being occupied by osteoblasts. These osteoblasts, once formed, have the power of direct and vigorous proliferation, showing great vegetative capa city; are capable of dissemination, of growing in the midst of the soft tis sue, or of being carried by the blood stream and deposited in the blood clot, where they proliferate after the matrix has been supplied with new formed blood vessels. As long as the bone cell remains embryonic, it ex hibits the power of proliferation; but when it reaches maturity it assumes the fixed tissue type and becomes stationary. This period is coincident with calcareous depostion and with it the cessation of active regeneration, 8.

(4) -426-. though its proliferating potentiality still remains. The diaphyseal bone is reproduced by the proliferation of osteoblasts derived from pre-existing os seous tissue and its regeneration takes place independently. of the perios. teum. Gedden7 1912.. He agreed that the periosteum takes no part in the. work of bone formation.. He, however, opposed that the osteoblasts was. derived from the cartilage cell, that cartilage was always present potential ly in bone even when mature and that when repair took place the deve lopment of cartilage preceeded the formation of bone. the osteoblasts are ectodermal in origin. his opinion. 1.. He gave two definite reasons for. If cartilage, in which ossification is just commencing, is sectioned and stained. with. which the. lay. Sabin's. down. osteoblasts. bright. the. which. cells.. chondral. osteoblasts. This. are. the. lineage. have,. for a time. it is always definite most. advanced. Paterson8. peared. from. 1912.. He. connective. occurred. Wm. one. osteoblasts. to be essentially. embryonic which. paths. trace. in the the. irritation.. bone than from cartilage,. membrane. bones. to remove. the periosteum. of the cranium,. the periosteum;. have. to the tint. that. any. that. and. continuous. and same. of the. the. endo and. direct. it. genetic. cells of a common. no sections. chain. numerous. of. points. are missed,. osteoblasts. and. of irruption. to the. cartilage. fact. that. the. He thought. He pointed. erupt,. subperiosteal,. cause. that. out that. there. and he doubted bone. that its future behaviors. agent. ap. was. was a chemical. an one. was extravascular.. as evidenced. from a healthy. of ossification. the. tissue cell and that the process. 1912.. osteoblasts. reaction.. in situ, and that the ossification. Wright9. stained blue-red. the. they. sectioned a. of forming. fecting. staining. or the. upheld. from. for it is rule,. similar. to. all the. improbable. that. is serially. possible. cell. things. the. the cells which. from. it extremely. cells;. stain,. bone. markedly. different. cartilage. examined. of bone,. improbable. with. Mallory's. endochondral. makes. it extremely. tissue. rind. differs. relation. If the. of. peripheral. forming. red tint,. makes. modification. the. cartilage. also. 2.. He believed that. must. be other. in the formation whether. without. ways of the. it was possible fundamentally. could scarecely. af. be regard 4.

(5) -427-. ed as the normal behavior of intact periosteum.. He further believed that. the evidence of comperative anatomy was in support of Gedden's conten tion that bone was ectodermal in origin. Douglas-Crowford10 1912.. They accepted Macewen's view of forma. tion and growth of bone. Mayer and Wehner11 1914.. They opposed the Macewen's opinion and. both agreed on the point that periosteum transplanted in the muscle of a young dog and periosteum after subperiosteal bone resection on the rib of a young rabbit produce new bone. There was, however, no bone forma tion in the "capsule test". by covering the bone with steel or glass, where. the periosteum was removed.. They also observed that in the bone trans. plantation, the bone cell does not show its activity, but the periosteum, as well as the endosteum has the function of bone formation. Gallie and Robertson12 1914. They viewed that the periosteum is not osteogenetic, and the presence or abscence of the periosteum in autoge neous bone transplants has no decided influence on the activity of the sub periosteal osteoblasts, however, it is of great importance, because of its control of the circulation throughout living bone. When a piece of bone has been cut free from its circulation and transplanted. into other part in. the same animal, if its surface is freely bathed in lymph, those cells which are present on the surface and in the open mouths of the Haversian's canals will live and undergo rapid proliferation. The cells in the lacunae or those deep in the Haversian's canal die, and in the course of two or three weeks disappear. The proliferating cells on the surfaces, whether derived from the graft or from the neighboring living bone immediately commence the absorption of the graft, and do not cease their activity until every vestige of the old bone has disappeared.. Bone transplanted. from one animal to. another of the same species may act similarly to autogeneous transplants. It probably always does so in dogs and cats, but in man there is reason able doubt as to whether the cells from all bones will live in the fluid of all recipient. Minoura13 1914.. He transplanted the metatarsophalangeal joints, either. intact or split longitudinally, of two months old rabbits, into subcutaneous tissue of the back, into the liver, or in abdominal cavity of the same or 5.

(6) -428-. different animals. The joint cartilage remained intact for a considerable time and always showed a cartilage cell regeneration on the periphery. The marrow degenerated and was substituted by fibrous tissue and later by fat. The trabeculae of the epiphysis degenerated and later was substituted by new formed bone, which however was not permanent. The epiphyseal cartilage line underwent necrosis and absorption, only the parts of the pro liferating zone near the periphery showed multiplication of the cartilage cells. The periosteum always showed new bone formation in the inner layers, but the new formed tissue finally underwent absorption. Nakahara14 1904. He injected the periosteum of rabbit, which was smashed to pieces, in the subcutaneous-and muscle-tissue and he observed from the transplanted periosteum, the newly formed bone, cartilage and also a transitinal type from osteoid to real bone tissue. Goto151914, and Tongu161915. They also proved that the transplan tation of an emulsion of periosteum can form new bone. Yokoi171912. He described the like result as above one. He added that when it is injected an emulsion of periosteum with fresh blood, he could not prove any special increased bone formation, but when one per cent solution of fibrin is substituted for blood there is apparently increased proliferation on new bone. Tsunoda181912. He observed that an autotransplanted periosteum in the muscle tissue of a young rabbit has an activity to grow new bone. He succeeded the above experiments in six from 44 cases (15%) within two weeks after transplantation, however, failed of the transplantation of the so called periosteal osteogenetic cells, which were romoved by scrubbing the inner surface of the periosteum. Goto151914. He said that the transplantation of an emulsion of so called subperiosteal osteogenetic layer in normal saline solution failed to grow new bone, on the contrary, the direct transplantation of the above one without the saline solution, following Pochhammer, can grow new bone. Murphy491912. He concluded that the periosteum fully detached from bone and (1) transplanted into a fatty or muscle tissue bed in the same individual, if he be young, may produce a lasting bone deposit; (2) trans planted into another individual or animal of the same species under the 6.

(7) -429-. same condition, it rarely produce a permanent bone deposit; (3) trans planted. into. Bone. with. individual,. and. sorbed, into. another. another. in the. into. absorbed.. Bone and. end. or both. the. living. bone. of the. asepsis. same. been. as when. had. the. scrapped, other. shape. Transplanted. dies. ultimately. and. in the. osteogenetic. for. the is. same. bone. becomes. the. as the. is ab. without. always. a scaffolding. if the. the. and. periosteum role. and. whether accidental surface.. in. at one. united. reproduction. transplanted. to. of new. fragment,. likely the. would. the. marrow. His. repair. presence. if. bone. its outer. sharp. to be deep. it is largely. elevater. occur.. on. an early. the and. It. bone more. a ques. is deep the. osteogenetic. with. bone On the. a blunt. to include. a connective. would. and. compact. the. as to whether. the cleavage. enough. of the. periosteum. on which. it includes. off or remooved. periphry. in. the os. bone.. In. that. limiting. membrane. is believed. that. graft,. play. abundant. the. an. impor. blood. supply. transplant. theory All. is that the. of fracture. of the. that. to be actively. substance. to the. a. the. grafts. If by chance. constitute. to establish. of bone.. Living. sure. which. to bone. it seemed. is stripped. on. from. is and what. with. or no osteogenesis. 1914.. in the. from. is very. is not. bone. mother. also. or not.. periosteum. in aiding. Graves21. and. is removed. of cells. bone. satisfactory. periosteum. osteogenetic. periosteum. recipient. the. the. cleavage. slight. that. equally. retained. of what. layer. instance. considered. periosteum. the. teogenetic. 7. same. periosteum. hand,. mainly. and. been. the. strument,. not. size. proved. it is to be actively. from. as. He. removed. tion of definition. tant. acts. 1913.. periosteum. only. and. fragment. and. transplanted. always. living. same. is obtained.. Albee20 had. transplanted. dies. transplanted. other. deposit.. in the. or infants.. Bone. tissue. bone. etc.,. always. children. periosteum with. fat,. practically. or cellular. in contact. fragment. a permanent. muscle,. absorbed.. or without. of the. into. of very young. muscle. with. coming. produce. contact,. it is always. the. ends. bony. case. species. periosteum. it never. transplanted. free from. except. dividual. species. its periosteum. outer. is the. chief. or periosteal. the. periosteum. osteogenetic. properties. or in grafting, layer. of bone. source. and. surfaces,. is the product,. of the periosteum,. are due to the more cells origin and. and. or less,. adherent. to its deeper. of callus,. which. to a less. extent. grows from. its.

(8) -430-. deep or medullary surfaces and its cut ends. Fujimori221915. He viewed from his experiment that the fate of the periosteum, whether it is auto-, homo-, or hetero-transplant, or it is either old or young, when it is transplanted in the subcutaneous or muscle tissue and quite free separated from the bone system, always undergoes to the regressive degeneration. From his 40 cases of experiment's he could find none of bone formation but a case of cartilaginous tissue. Tomita231908. He noticed that the bone cell has no activity in bone formation, but it grows always from the periosteum and marrow cells. He also proved that a piece of bone transplanted into the abdominal cavity kept its vitality for 95 days. Ryerson241913. He illustrated by X-ray examination that the bone regeneration appeared in two cases in which the fracture occured in infancy, and the X-ray showed a shadow a quarter of an inch wide beneath the stripped periosteum of the femur and extending well into the osteo-perio steal angle. This fact is explained because at this age a rapid growth of bone occurs from the large number of osteoblasts which can be demon strated histologically beneath the periosteum. If periosteum were osteo genetic, then new bone would be produces beneath it over the area where it is stripped from the bone and the osteo-periosteal angle would be one of the first parts to be filled. That this is not the case is manifested by the plates which show that the osteo-periosteal angle is actually the last part of the space to be filled with a shadow, indicating new bone forma tion. From this, therefore, the natural inference is that the periosteum is not osteogenetic in character. Bancroft251914. From his experiments, two types of repair of bone have been observed. 1. Where the periosteum has been well stripped off the bone for a distance of several centimeters it is separated from it by serosanguinous fluid, here the bone has apparently been repaired from the medullary cavity by a framework of connective tissue upon which calcium salts have been deposited, forming osteoid tissue; there has also been a reparative pro cess from the connective tissue of the Haversian's canals at the sides of the bone incision. 8.

(9) -4312.. Where. the. capsule,. bone. muscles.. other. the. in shape. and. serve. Davison. structure. the. "periosseous. genetic. layer. densation tion the. presence. genetic. osseous. to some. that. can. They. are. together. cells. with. in the. for bone. periosteum. is the. and. the. retention. corresponding. and. of the. word.. form. same bone,. may. of vitality intercellular. that that. neither neither. He. also. osteo upon. canal. graft,. bone. The. most. matrix.. "formes fibrous. fibrous. tissue. if there. other. osteoblasts.. os. implanted. nor marrow. as any. enough. from the osteo. calcium. as well. i.. the. large. of the. as other. into. implanted. in some. the. the. of the bone,. obtained. of them that. without. periosseous. of the. periosteum. said. a con. surrounding. medullary. be. osteo. of differentiation. of an osseous. as. This. Regenera. structure. developement. stimnlus. consti. is dependent. entire. the. cellular. undergoes. of the. and. is called. be accomplished. especially. about. propers. ........... capable. has. cellular. layer. bone.. mesenchym periosteum. the. continued. such. produce. cells,. in the transplantation. formation,. is the will. layers. cells. to con. very. of bone. of its. osteogenesis. layers,. is. the periosteum. cellular. virtue. components. throughout. there. tends. is found. cannot. into bone. He described. meaning. by. the. that. osteogenetic. their. Ely27 1919.. 9. scattered. with. host. noticed tissue. purposes,. of the. it, they. also. cellular. that. This. periosteum. of the. are larger. osteogenetic. to the bone,. is a part. the surrounding. transplanted. to. supply.. and. .......... different. In places. cells to these. to proliferation,. before. be expected. teoblasts,. proper. since. membrane. extent. for practical. sary. layer". periosseous. and. bone. capsules.. of connective. e., in the. blasts. cells. not. approximation. stain.. periosteum. small. a staut. tissue,. the. tissue. to the sur. of cells which. summarized. tissue. of some. maturation. They. stimulated. layer. membrane. Beneath. is present. layer.. tissue. its blood. 1916.. adherent. in immediate. a heavy. fibrous. when. from. take. to furnish. osteogetic. into. of bone. that. of many. overlying. it can,. that. also. connective. is apparently. is a single. connective. Smith26. composes. of the. tuents. there. properties.. not. except. fibrous. and. tissue. as a limiting. and. and. osteogenetic. the. acts. its shape. periosteum. tissue,. nuclei. from. periosteum. a definite. connective. bone,. have. a metaplasia. no. This. forming. under. with. connective. newly. The. repaired. it is continuous. rounding from. has. is neces. bone," tissue. is osteoblasts The. in the of the under. osteoblast.

(10) -432-. is the bone building cell, if the present theory of bone formation, the neo plastic theory, is correct. The function of bone needs three important conditions as follows: 1. Blood vessels. 2. Either a loose meshed fibrous tissue, or a homogeneous (cartilage matrix 3.. A. or a granulus. stimulus,. Hyde28 osteum,. He. such. has. of regeneration. not. a necrotic. physiologically. 1920.. as. or. or pathologically. classified. chiefly. 1885.. He. viewed. loss. vitality. but. multiply.. Bart30. 1921.. osteoblasts. but. in some. way. He. stated. related. to bone. bone. cells. that. cortical. bone.. are. that. whether. new bone or not.. difficult. to determine. with certainty. is formed. bing lightly. some amount. the boundary. examination. the cut-edge. Even. great. on under. ether. which. are. has the. mentioned, periosteum. is present. as the same.. must be always. of technic. experiments. To cut. of. is made,. the periosteum. required.. and. and scrub. knife, a delicate flap. of thus obtained. very. before the. with forceps,. delicacy. from a portion. and some cat and dog were used for these. The tissues. of the. structure. of the femur underneath. and therefore,. cells. chisel was used and made a rectangle. pulling. off.. of cortices. do. functionless. the periosteum. of the bone with a sharp. is stripped. times detached a microscopic. a sharp. the surface. the periosteum. active. It is as above. the osteogenetic. By pinching,. transplant. senile,. and so they must not be considered. the periosteum,. on the femur.. in the. not. are. of all, it was investigated. to proliferate. operated. of the. Materials.. periosteum. peri. the power. and. In the embryo,. that. 2, that. Methods. the bone.. -1,. be.. power,. cells. they. may. production.. activity. remove. layer. the. bone. indicate. III. First. that. that. observations. outer. case. as follows:. osteogenetic. in the. Bonome29 their. as the. his conclusion. no independent. resides. material.. is some periosteum. Mostly young rabbit and they were all. anesthesia.. were fixed. in Zenker's fluid. or 10 per cent. solution. of 10.

(11) 11. -433formaldehyde, then washed. decalcified thoroughly. in 5 per cent solution. of nitric. acid if necessary,. in one per cent solution of sodium carbonate. removal. from the nitric acid solution. loidin.. Stained. in Mallory's. eosin. and then embedded methylene. after. in paraffin or cel. blue or alum. hematoxylin. eosin.. IV.. Experiments.. Experiment I. The. autotransplantation. was made into the muscle outcomes. of periosteum tissue of back. of rabbit. mentioned. or cat.. The. 1.. 'a'. signifies the weight at the time of transplantation. ' b'. signifies the weight at the time of examination.. The. later,. or thigh. as above. are as follows:Table. amount. obtained. transplanted of fatty. was. tissue. ultimately. periosteum. disappeared. substituted. the. absorbed. and. .. in a short. locality.. none. of bone. Some. time. underwent. formation. could. and. some. necrosis, be found..

(12) -434-. Appendix of Experiment. 1.. On Nos. 19C and 21C, ca. 1 c.m. by 0.2 c.m. flap "of periosteum was stripped off, with attaching. one end to the femur and this pedunculated. periosteum was sutured to an adjacent muscle tissue. After 5 weeks and 32 days respectively, it was observed that thus pedunculated periosteum has changed into a bony spicule and at its top, a silk-gut suture previously used was discovered.. This experiment is quite different from that of Ex. periment 1; viz., the former, the periosteum is entirely detached and trans planted freely into muscle tissue where the bone does not physiologically exist and the latter, it is a pedunculated periosteum attached at one end to the femur and so it is a continuity from the thigh bone. Experiment. 2.. A composition cium normal. phosphate saline. of magnesium. as following solution. phosphate,. proportions. was made,. calcium. was prepared. then transplanted. carbonate,. and cal. and an emulsion in. into the muscle tissue. of rabbit.. Composition of Human bone. Mg3(PO4)2.............................. 1.75 CaCO3.................................... 9.2 Ca3(PO4)2 .................................87.8 by Carnot.31 Microscopic Examination: I No. 26B 5 weeks after transplantation. There are several particles of artificial bone calcium in various sizes in the transplanted bed and the smaller one was fused with so-called for eign-body giant cells and the bigger one is capsulated with some layers of newly formed connective tissue cells and its inside was invaded with the same cell-groups. Several newly formed blood vessels also occupied the locality. In some place, the calcium particles nearly disappeared, having left a group of giant cells. The artificial bone calcium was also transplanted in the subcutaneous tissue of dog, rat and mouse and examined in various periods. Still, the injection of the bone calcium emulsion as above mentioned, was made once every day continuously for 25 days into the muscle tissue of dog and rab 12.

(13) -435-. bit. Still more, to give continuous stimulation in the injected portion, the artificial bone calcium emulsion in two per cent solution of formaldehyde was used. This 2 per cent solution of formaldehyde gives good result on tissue for continuous stimulant but does not destroy the tissue. All the tests that were attempted failed for bone formation, and the results were practically similar to that of No. 26B on the main points. Experiment 3. The bone dust was substituted for the above artificial bone calcium, and the bone dust was made as follows:Material: rabbit's thigh bone. A thigh bone obtained from a young rabbit was preserved into 10 per cent solution of formaldehyde for a few days and then boiled in water for an hour. Both the periosteum and the marrow were entirely removed, then the minute bone dust was made and sterilized. It will be summarized the results of 7 cases in rabbits and cat, extended from 9 to 131 days. The bone dust was transplanted into sub cutaneous or muscle tissue as previous cases, however, all these tests failed to form new bone. The transplanted portions were generally occupied with fi brous and fatty tissues and there formed many so-called foreign body giant cells around the bony particles. In later period, the bony particles have become nearly homogeneous or disappeared, having left several groups of giant cells. In early period, there were found some unaffected bone dust and some extend of hemorrhage in the transplanted bed. Experiment 4. By Experiment 1, it is affirmed that the periosteum itself transplanted freely into the muscle tissue has no activity to proliferate new bone, Here, the transplantation of the periosteum intermingled with above bone dust was attemped. Method: -A flap of periosteum removed, with above mentioned-atten tion, from the femur of rabbit was intermingled with the bone dust and transplanted in the muscle tissue of the loin or the thigh of the same ani mal. The results are shown in Table 2. 13.

(14) Table. Notice:. 2.. Homo-transplantation was tried in Nos. 21 to 25, viz., a flap of periosteum was obtained from a young rabbit and transplanted in the muscle tissue of another young rabbit of same species..

(15) -437-. There osteoid.. could. be obtained. However,. velopment. all of the positive. of new bone.. will be here described No. 55A.. 5 positive. results. from. 26 cases. and. one. cases but one showed very poor de. No. 55A only showed. an excellent. result. and. in detail.. 10 weeks after transplantation.. Gross finding: - All the wounds operated able appeared. in the transplanted. only a guidance. in finding. lump was palpated Microscopic. on were completely portion. and. the. black. remark. silk suture. was. A hard. on the spot.. examination:. - The transplanted. bed was occupied In the connective. formed. which appeared. bony tissue. a grain sized mass.. This young. and somewhat flattened whole. bone.. many. red blood. with abundant fibrous. tissue,. there. macroscopically bony. There also appeared corpuscles. young. some Haversian's. and fibrous. with several. layers. periosteum.. There. sizes around. the. homogeneous. and some nearly. No connections. were also many. bone dust.. between. absorbed,. the new. body. particles. bone. the. smaller than osteo This new bone was. giant. cells, i.e.,. the. cells in various. of bone dust. having. lined. canals which contained. of many flattened. foreign. These. many well stained. osteoblasts. cells, somewhat. consisted. and. of newly. as an oval shaped and. tissue possessed. bone cells and large. tissue. was an area. and its inside was also lined with osteoblasts.. surrounded. Nothing. the locality where it was transplanted.. some fatty tissue.. blasts,. healed.. were mostly. left a group of giant cells.. and the giant. cells seemed. to be. existed. On negative some. cases, generally. fatty tissue occupied. particles nective. in various. the. sizes could. tissue cells seemed. the. newly. spot and. formed. giant. be seen in situ.. to have. connective. cells fused the. tissue. and. bone dust. These newly formed con. come from that. of the host's. muscle. tissue. For comparison,. one half of the. removed. periosteum. used in this ex. periment was always transplanted without bone dust in the muscle of the opposite side and none of bone formation could be found.. 15. tissue.

(16) 16. -438-. Ex periment 5. Periosteum the. muscle. tissue. sprincled of young. with. artificial. rabbit. Table. bone. The. Table. calcium. was. 3 illustrates. transplanted. in. it.. 3.. There showed new bone formation one in 7 cases, and the size, shape, and ist appearance of the new bone is quite similar to that of No. 55A. It was situated in the connective tissue of the host's muscle bed and no fatty tissue was existed. No particle of artificial bone calcium nor giant cells could be found. Experiment 6. A piece of bone removed from the femur of a rabbit was completely denuded its periosteum, subperiosteal layers and endosteum and transplanted into muscle tissue of the same animal. On the other hand, a piece of bone which possesses completely its periosteum as well as endosteum was re moved from the femur of a rabbit and transplanted into muscle tissue of the same animal. Microscopic Examination: - The former transplant, at the seventh week, the bone cells of No. 6 poorly stained and some disappeared. The periphery of the transplant was irregularly and extremely eroded and was surrounded by newly formed large cuboidal cells at the adjacent, and still by slight flattened cells. There.

(17) -439-. was no newly formed bony tissue found. At the tenth week, in No. 5 case, the regressive process still further advanced and no more stained bone cells could be found.. Groups of fibrous and fatty tissues were only left. on the transplanted portion. The latter transplants Nos. 15 and 16, showed both at the fifth. and. tenth weeks, their bone cells stained fairly well, and moreover, present new ly formed bony tissue which stained light pink more than the old bone. However, the proliferative activity in both cases has been suspended. Furthermore a patella was buried into muscle tissue of the another rabbit in two cases, Nos. 55C and 56C.. After 10 weeks and 80 days re. spectively, in both cases, it was discovered that the bone cells had either disappeared or strongly atrophied but the cartilage cells situated at the junc tion where the bone part transfers to the tendon have stained fairly well. Some of the Haversian's canals were filled with vascularized fibrous tissue, the other contained many polymorphonuclear cells and red blood corpusc les or the latter only. Still more, the red blood corpuscles were nearly homogeneous and some showed fibrin formation in several places. Besides these cases, transplantation of tail-bone of rat and mouse was attempted and this result will be reported later by another paper.. Experiment 7. In this test, the surface of the femur of a young rabbit was strongly curretted and accordingly the periosteum thus obtained unquestionably con tains some amount of subperiosteal layer and cortices of femur. Such a fl ap of periosteum was transplanted into muscle tissue of the same animal and the result is shown in Table 4.. (Table 4 s. next page.) Microscopic. Examination:-. By this examination, which was surrounded. 3 in 12 cases showed. newly. with large cuboidal or somewhat. bone tissue. flattened. are seen some. particles. their bone cells.. Several. of giant cells are found around these dead. bone particles 17. bone. osteoblasts.. In No. 31, there. groups. of dead. formed. and some of them were nearly homogeneous.. which. have. lost. These groups.

(18) -440 - Table. of active and non-active layers showed fi brous. of connective. bony tissues are surrounded,. tissue cells in the muscle. two groups of new-bone-islands tissue.. the fibrous. 4.. In addition,. for a whole, by several. bed of the host.. No. 81B. which were thickly surrounded. there are seen several. groups. of giant. with cells in. tissue.. Appendix:After. removed. a flap. of periosteum. the inner surface. of thus removed. slightly. and a coagulated. curretted. ly, it is so-called. subperiosteal. from the femur of a young rabbit,. periosteum. and the denuded. bloody-like. osteogenetic. femur were. mass was collected;. layer.. This mass was. name trans. planted into muscle tissue of the same animal. This was tried only in one case No. 49C and after 10. wooks, no trace could be found in situ. Experiment. 8.. On this test, the transplant,. similar to that of previous. case. was in 18.

(19) - 441termingled. with. periosteum, bone. bone. dust,. subperiosteal. dust. result. layer,. mentioned,. some. amount. of cortices. into. muscle. tissue. shows. 5.. on. Table. Table. 2 in 4 cases showed an excellent. namely,. autotransplanted. were. The. as above. proliferative. No. 56A.. 80 days. positive. the. mixtures. of the. of young. femur. of and. rabbit.. 5.. results,. and especially,. No. 56A showed. activity. after transplantation.. Gross finding: - The denuded which had suture. portion on the femur was slightly. been made on it could be found except. there is seen black silk. Examination:. - In the section,. having. and a hole. on the spot.. Microscopic. which. protruded. there. can be discovered stemmed. appeared. newly formed bony tissue in two groups. by naked eye.. with connective. They were situated. tissue which. have. in fatty tissue,. arised from the host's. muscle tissue and these two groups of bone were also connected with connective. tissue band.. In both groups of the bone, some bone marrow. has been formed and they consisted of red-blood-corpuscles cuboidal. each other. cells, viz., the marrow. cells.. No. and well stained. bone dust could. be found. in. this case.. Experiment. 9.. It is well known that the endosteum periosteum has the 19. and therefore, activity. has the same function. it was here investigated. to proliferate. new. bone or not.. whether. with the. the endosteum. It is more. difficult. to.

(20) -442-. determine, with certainty, the boundary of the endosteum and the bone. It is practically impossible to remove the endosteum alone and when it is removed, not only, the endosteum but the bone marrow always accompany to the endosteum. Furthermore, the inner surface of the bone in contact with the endosteum is rather softer in hardness than that of the bone in contact with the periosteum and this fact stillmore increases the above dif fi culty. On this test, the transplant, therefore, consists of endosteum, subendo steal layer, some amount of bone part and bone marrow. The result is shown in Table 6. Table 6.. As the table shows, the percentage of the bone regeneration. is quite. large and the proliferative activity is very marked in all positive cases. Several cases will be here described in detail. No. 19B. b weeks after transplantation. Microscopic Examination: -In the fatty tissue of the transplanted. bed of the host, there are seen. two groups of newly formed bony tissues and their appearances like a coral reef in the sea of fatty tissue.. These circled islands of new. bone hold some amount of marrow rich in red-blood-corpuscles outer layer is surrounded with fibrous tissue.. are just and their. The bone cell's as well as 20.

(21) -443-. the osteoblasts are quite larger than those of surrounded connective tissue. Generally, at fifth to seventh week after transplantation, tive activity is in its maximum and later it is gradually. the prolifera. going to regres. sive degeneration though the newly formed bone still retains its vitality at tenth or twelfth week. At tenth week, No. 10B showed a small mass of new bone in the transplanted bed, having surrounded with connective tissue, and No. 20B also showed the similar condition in the fatty tissue of the host.. Commonly, the coral reef appearance of newly formed bone seems. to be a special feature in this experiment. Appendix: - Furthermore,. the transplant as previous case, with either bone dust or. artificial bone calcium was used but so satisfactory result was not obtain ed by adding bone dust or artificial bone calcium.. The former showed. one positive in 4 cases and the latter, 2 in 3 cases. cases will be stated.. Some of the positive. Microscopic Examination: - No. 14A.. 5 weeks after transplantation.. This appearance is quite similar to that of previous one No. 19B, ex cept there are several small groups of giant cells which have fused the homo geneous bone dust particles. No. 19A. 5 weeks after transplantation. A fairly large area of newly formed bony tissue which is very irre gularly constructed a lattice was situated in the fatty tissue. No bone marrow could be found. The particles of artificial bone calcium nearly dis appeared. No. 20A.. 10 weeks after transplantation.. There are two small areas of newly formed bone which are surrounded with fibrous tissue in the fatty tissue and its proliferative activity is very faint, compared with the previous one. Several groups of giant cells fused the particles of artificial bone calcium which are nearly disappearing were also found in the fatty tissue. Experiment 10. If the fact that the endosteum has the same function and is similar 21.

(22) -444-. tissue. with. from. where. with. Experiment. marrow row. the the. from. was. layer,. some. activity. is consented,. marrow. 7.. was. However,. to the. compared. almost. the. No.. 60A. Microscopic - No. A. especially. 60A.. planted muscle -osteoblasts-and. sign. revealed. and. the. so the bone. consists. bone mar. of periosteal. marrow.. It revealed. Experiment. Table. 9. to compare. to exclude. test. bone. of Experiment. 9,. both. the. the proli. 7 is illustrated.. 7.. a great. deal of newly. formed. bone. occupied. the. tissue.. Examination:. quite. shaped. displayed. this. with. Table. be. mentioned,. and. same.. result. must. 7, viz., of femur. the. impossible. above. Experiment. having. were. excluded. 9, as. of cortices. and. then. it is quite. Experiment. amount. result. ferative. bone. the. added. excellent. periosteum. cells.. 5 weeks large bed. No. area. bone. transplantation.. of newly. formed. bony. tissue. trans. of the host and it was lined with large cuboidal cells; was still surrounded with several layers of large spindle fatty. tissue. of "Halisteresis". homogeneous. after. was dust. around observed. was. the. new. in some. obtained. bone spots.. in the fibrous. could A. be found. few. The. particles. of. tissue.. Appendix: - The plant. Experiment. as in Experiment. 8 showed 8. was. positive mixed. results with. and. bone. here marrow. the similar which. trans. obtained 22.

(23) -445-. from the femur of the same animal.. The. periosteum,. amount. subperiosteal. bone marrow. transplantation. Several. groups. layer, some. No. 53A revealed The. transplant,. a positive. new bone was situated. of foreign body. giant. therefore,. consists of. of bone part, bone dust and result. at the fifth. week. after. in the fatty tissue of the host.. cells which. consist. of homogeneous. bone dust were also observed. Experiment 11. The bone marrow rabbit. and transplanted. was obtained. was selected. from two parts. in the muscle. from the central. part. tissue. of a femur. of the same animal;. of the femur and. the other,. of young the one from the. periphery to come in contact with the endosteum of the same femur, and the results between the two were vice versa. Table 8, A and B show the result.. The former result. is shown in Table. 8 B.. 23. Table. 8. A.. Table. 8. B.. 8 A and the latter,. in Table.

(24) - 446-. The former was investigated in six cases and failed to form new bone in all, but the latter revealed three positive ones from six cases and its ap pearance was quite similar to that of Experiment 9, that is, the new bone was distinguished along the endosteum and the bone marrow also exhibited their proliferative activity. Experiment 12. Either the bone dust or artificial bone calcium was mixed with the bone marrow of young rabbit and buried into muscle tissue of the same animal and examined after 5 and 12 weeks respectively.. However, no. newly formed bony tissue could be found and the locality was always oc cupied with fatty tissue.. Foreign body giant cells also fused the particles. of either bone dust or artificial bone calcium. Experiment. 13.. In this test,. the bone. viz., the transplantation attempted. No. 62A.. microscopic. bone. was obtained. to that dust and. of Experiment bone. marrow. from four cases. 4; was. in which. in two cases.. after transplantation.. appearace. so will not here describe. bone calcium. result. was homotransplant 5 weeks. Furthermore,. was added. of periosteum,. and one positive. the bone marrow. The. marrow. was quite. similar to that of No. 19A and. it.. the transplantation. and bone marrow. which consists of periosteum,. of the same animal was tried. artificial. and was be. able to form new bone. No. 68A. There. 10 weeks. is an irregular,. after transplantation. nearly. homogeneous. group of artificial bone cal. cium in the fatty tissue of the host and it is capsulated On this spot, a semicircular with large. and somewhat. ered with several. new bone was situated, spindle. shaped. layers of flattened. cells.. The. spindle shaped. with fibrous. tissue.. having been mediated free surface was cov cells.. Experiment 14. The. spleen. has a resemble. function. with the bone marrow. and so in 24.

(25) -447this test, the spleen. emulsion. of Experiment. An emulsion. 10.. was aseptically fi ced.. having. rabbit mixed. was. was prepared. for the bone marrow. from a piece of spleen which. from a young rabbit immediately. A piece of periosteum. a young. result. obtained. in fresh was substituted. removed. transplanted. with the. above. by strongly. into muscle prepared. curretting. tissue. spleen. after it was sacri the femur of. of the same. emulsion.. The. was obtained.. Experiment. 15.. The spleen. emulsion. was also substituted. for the bone marrow. periment 13. The transplant, therefore, consists of periosteum, and spleen-emulsion. The result was also negative. Both the above two experiments must be further. more investigated. spleen emulsion. in this occasion,. Summarizing at. the. above. must be noticed. and so that. the. was a homotransplant.. Summary.. experiments,. the. following. table. shows. the all. Total N os.. Positive Nos.. a glance.. Nos.. Kind of transplant.. Result.. 1. 2.. Periosteum (free) ... ... Periosteum (pedunculated). ... .... ... negative ... positive. 10. 2.. 0. 2.. 3.. Artificial. 4.. Bone dust, .... 12.. 0.. 7.. 5.. Periosteum. plus bone dust,. 0.. 26.. 6.. 6.. Periosteum. plus artificial. bone calcium, ... ". 7.. 1.. 7.. Denuded. bone, .... .... .... .... .... ... negative. 2.. 0.. 8.. Complete. bone, .... .... .... .... .... ... positive. 2.. 9.. Periosteum,. 2.. ... ". 12.. 3.. layer, ". 1.. 0.. bone calcium, .... .... subperiosteal. mount of bone-part, 10.. So-called. 11.. Periosteum,. 25. in Ex. bone dust. were limited to a few cases. and stillmore. V.. results. animal, negative. subperiosteal subperiosteal. ... .... .... .... .... ... negative. .... .... .... ... ". .... .... ... positive. layer, .... some. .... .... osteogenetic layer,. some. a. a.

(26) -448-. Nos.. Kind of transplant. mount. 12.. of bone-part. Endosteum,. of bone-part,. Endosteum,. 14.. dust,. Endosteum,. mount. bone. 23. 24.. bone .... .... of bone-part. Periosteum,. bone .... .... layer, bone. .... .... Positive Nos.. 4.. 2.. 8.. 7.. 4.. 1.. 3.. 2.. 8.. 5.. 1.. 1.. 6. 6. 2. 2. 4.. 0. 3. 0. 0. 1.. 1.. 1.. 3. 2.. 0. 0.. a. ... some. ... " a plus. ... some. ... " a. marrow, " some. a plus. .... ... ". Bone marrow (central part) ... ... ... negative Bone marrow (periphery) ... ... ... ... positive Bone marrow plus bone dust, ... ... ... negative Bone marrow plus artificial bone calcium, " Periosteum, bone dust plus bone marrow, positive Periosteum, artificial bone calcium plus bone marrow, ... ... ... ... ... ... " Periosteum, subperiosteal layer, some a mount of bone-part plus spleen, ... ... negative Periosteum, bone dust plus spleen, ... ... ". VI.. Total N os.. plus. marrow. .... positive. ... ". some. .... bone. subperiosteal. dust,. a. marrow. layer,. plus. of bone-part,. some. .... layer,. calcium,. .... marrow. subperiosteal. mount. 17. 18. 19. 20. 21. 22.. layer,. of bone-part,. Periosteum,. bone. .... dust,. marrow,. subendosteal. artificial. 16.. layer, bone. of bone-part. mount. 15.. bone. subendosteal. mount bone. plus. subendosteal. mount 13.. Result.. Appendix.. For an appendix, a few lines on bone formation which occurs without any relations with bone-system will be here added.. There are reported. bone-formation in almost all the organs and tissues and such instances are not so infrequent.. Several instances which hitherto have been experiment. ally proved will be here stated.. So-called myositis ossificans which grows 26.

(27) -449-. after operation or trauma in abdominal muscle tissue and etc., where there is physiologically no bone tissue existed is well known. The periosteal type must be excluded from this instance and it must come from the meta plasia of connective tissue of muscle (Goto15,Nakayama38,etc.). Bone for mation in the artery, especially in tunica media of full grown rabbit with a 3 per cent solution of silver nitrate or a 2 per cent solution of cupric sulphate application was proved by Harvey39,etc. By ligation of renal blood vessels on rabbit, some bone tissue can form in the affected kidney (Liek40,Koga33, Asami32,etc.). Furthermore, bone formation in the tonsil, thyroid, As. ovary,. above. where. liver, heart,. mentioned,. there. periosteum An. lung,. the. bone. is absolutely nor. bone. bone-system concerns. onn renal. Animal. No.. Extra. Weight. 2625gm.. vessel. 1.. existed,. on. frequently. independently. bone. ligation. Species. Sex. are hitherto,. can be formed. no. marrow. experiment. eye, etc.,. reported.. in such. and. a place. consequently. no. formation.. of a rabbit. was. attempted.. renal. vessels. rabbit.. male. Age. 1 year. old.. Method: - Opened silk. gut. the. after. Sacrificed. right. the. after. side. of abdomen. suprarenal. 10. capsule. weeks.. and was. Weighted. ligated. the. stripped. 2362gm.. off.. Closed. with. the wound.. on examination.. Gross finding:-. The right kidney has been buried in the fatty tissue and a part was strongly adhered to the liver. The shape was well kept but its size was nearly three times smaller than that of healthy side, and its consistence was just like cartilage. The cut-surface showed that it is occupied with white calcified masses, radiated from pyramid to cortex. Microscopic. Examination:. - Mostly with. calcified. arranged. fi brous 27. tissue. portion stained. directions. masses connective and there. and. some. Some and. the. tissue. were. medullary. homogeneously. of tubules.. disappeared.. calcified. vascularized. cortical. masses,. to the. apparatuses above. the. surface The. many. in pink,. Malpighians. amount. of fibrous of cortex. medullary groups. portion which. bodies tissue. of somewhat. occupied. radiated,. and. and all tubules. existed. is also portion. are. among. surrounded. mostly large. with. consisted. spindle. the. of. shaped.

(28) -450-. cells which. stained. in deeper. blue than. pelvis also consisted of fibrous tissue some bone marrow rich in red-blood activity. of this newly. - boidal. formed bony. bone cells were poorly. are many. muscle tissue periosteum. regenerates. that. This is the point subperiosteal. poor and the large cu. Discussion. the periosteum However, described Even. transplanted. periosteum. into. it is very difficult "to remove the. alone was really removed. if an attention. layer and still some amount. off with the. freely. and in such a positive case, it is. the periosteum. of discussion.. osteogenetic. times shaved. part. bone.. whether. The. and a bone tissue which possesses corpuscles appeared in situ. The. tissue was very. that. alone as previously. very doubtful. cortices. reports. tissue ground.. stained.. VII. There. that of fibrous. or not.. is paid, the so-called of bone part are some. on its removal.. Furthermore,. the. of the bone do not contain so much calcium than that of the deeper. and accordingly. the periphery. of the bone is rather. soft in hardness,. and especially it is still softer at young age. These matters increase the above difficulties on removal of the real periosteum. In the Experiment 1, none of positive cases could be given on the transplantation osteum.. of the peri. However, if the above attention is unpaid and when such a peri. osteum as it is removed by strong currettment is transplanted, relatively a majority of positive cases will be proved as Experiment 7 shows.. This is. the reason. that. auto still,. there. are many. or homo-transplantation added. removal plantation. etc.. why. that. of the and. the. periosteum so the. This fact relates. osteum. but the. amount. of bone part.. fact that. bone. bone. so-called. pro and con.. of periosteum pieces has cells. obviously. which lost. can. disappear,. Yokoi17 stated. be proved. were shaven. its vitality. within having. subperiosteal. periosteum. itself has activity. osteogenetic. to the appendix altered. new. bone.. off at the same a few days emptied. that he is transplanted. With regard. the pedunculated. if the periosteum. opinions. an He. time on. after trans. their. lacunae,. not only the peri. layer. and. still. of Experiment. some 1, the. into a bony spicule looks as. to form new bone,. However,. it can 28.

(29) -451-. not be adopted. By removal of periosteum, the so-called subperiosteal os teogenetic layer which was removed its protector the "periosteum" and the exposed bone part cooperate with the periosteum, thus it fulfils all condi tions required as that of Experiment pedunculated the. vessels,. to bone an. periosteum. blood. regeneration. important. to the. from. the. and. role. femur. femur.. the. 7. can This. periosteum. Moreover, the continuity of the be. supplied. matter may,. sufficient. nutrient;. is exceedingly. beneficial. on this ocassion,. rather. play. as a guide.. The transplantation. of either the artificial bone calcium or the bone. dust failed to form new bone.. Asami32 and Dock injected calcium salts. consisting of calcium chloride, calcium carbonate and sodium phosphate, subcutaneously or intramuscularly on rabbit, but no evidence of bone for mation was observed as late as 50days. after injection.. This experiment,. however, must be furthermore continued as far as the inetaplasia is gene rally consented. In fact, Koga33 proved new bone formation by repeated injection of one per cent solution of calcium chloride into subcutaneous muscle-tissue, or scrotum. could be observed. or. Goto15 also tried the same experiment and he. only an osteoid tissue.. fi rmed that the periosteum. By Experiment. itself has no activity. 1, it was af. to form bone, but the Ex. periment 4, viz., the periosteum combined with bone dust was favourable to the bone formation in a minority. The following things are great es sentials. gans The. to form new bone.. 1.. Early. and. 2.. Suitable. 3.. Non-pressure.. This. is out. and. tissues. transplant,. lar. tissue. which. the. stimulation,. The. locations. sufficient. establishment. of blood. supply.. stimulation.. of question. that. not. only. in the. require. early. and. sufficient. therefore,. must. not. be inserted. furnishes the. of the. muscle. an unfavorable motion. transplant,. transplant. blood into. environment. is an important. therefore,. cannot. or. supply;. the. cicatrical. or nonvascu. for it. agent. be. but all the. nutrient.. Referring. to. for this purpose.. disregarded. and. two. places were practically selected for comparison; the one is the muscle tis sue of both sides of lumbar column for the most quiet place and the other, the muscle tissue of the thigh 29. for the most movable. place.. However,. no.

(30) -452-. significance could be found between the two. It must be borne in mind that animals (rabbit in this occasion in the laboratory cages usually do not allow free excercise and so it may have no influence upon the loca lity. As to the favourable stimulant it will be described in the following pages. It may be considered that in such a place where there is no stimu lant the transplant will undergo to atrophia, later, necrosis, following the absorption.. Non-pressure. conditions.. The. the. upon. sternum. eloquently such. that. bone. muscle fiber. that. even. cannot. great. and. put. particles too. large. tively. requisite. other. rather. locality. time. undergo. and. bone. will. dust. to be. necrosis. absorption.. better. and. the. bone. age. and. nourishment. of animal,. the. bone. production.. It must. any. dust. a place. connection. tion hesitated. count. result. will must. where. be the. still. osteo-system,. Long. exposure. tive process.. As above. sion, requires. many. the. particle. are. that. much. Both. peri. this. matter. the. one. and. also. the. so it. will dust,. periosteum bone.. The. concern. for. rises freely. to exist,. makes. at. or. of the bone. in normal. allow. and reac. transplantation. not. bed. size of its. inflamed. of the. cause. does. was. to the location. other. proportion. to air and drying. enumerations,. It. It was,. if either. the. a predominal. time being prolonged. things. and. as they. and. along. in it.. is too. become. assimilate. wait. be considered. to the. gap. the. new bone.. and. The. it physiologically. of the operation. form. and. dust. having. smaller. similar. even. in the muscle. bust. If bone. be obtained.. long,. of the muscle and. a cavern. formation,. The. very. bone. in such a bed is easily pres. body. cannot. on. sternum. a transplant. of bone. be ripen. one. aneuryam. mass on examination.. to. bone. ripened,. the. in such. the. or. on. of requisite. a long. inserted. by making. interfere. must. the. pressure. quantity. for foreign. to cooperate. is delayed. and. conditions.. work. destroys. At first,. into a necrotic. The. breast. if it works. pressure. made. a good result. another. the. last. one. of the. pressure,. by the motion,. are. and. at. failure and the transplant. in it.. it will. same. was. a transplant. to the. osteum the. obtained. the. is also. pressure. a delicate. host. it had changed. afterward,. transplant. lasting. bear. sed out to the subcutane discovered. the. it impringes,. of the. was, however,. the. which. relates. a hard. the. fact. toword. the. without regenera. of the transplant. on ac. are also subsides the regenera. the bone regeneration. on this ocas. and it is very difficult to obtain the all requisits. 30.

(31) -453-. This is the reason why the positive cases are in quite small number. Whether the deposit of bone dust will work upon the proliferation as it stands or after it dissolves is the problem, but the latter will probably be adopted. b Koga33 viewed that on the ocassion of bone formation by injec ting one per cent solution of calcium chloride, an insoluble calcium which had. deposited. formed. in the. from. the. transplantation. previous. one.. This. the. dust. and. the. result.. of the. two. The. denuded. onset. when. planted cess,. into. tells the. bone. there. on. plantation. the. is successful From as the. the. endosteum. two. layers. is essential. than. the. that. differences. no. of. between. chemical. doubt,. period. com. necessary is likewise. it is poor,. of bone. tissue. It the. for bone. during. its pro. there. is no. on regeneration. and. as that of Experi week, the denuded. but at the tenth week,. both. the. Experiments that. formation.. the. the trans. that it is the com periosteum. is obvious. it is ascertained. at. and trans. On the contrary,. it contains. as both. vitality. rabbit. because. At the seventh. though. has. However,. degeneration,. or anything. 9, viz.,. bone,. results,. smaller. of a young. animal.. regressive. 7 and. new. On. one in 7 cases. the. bone piece is to be considered. all. to form above. 6,. bone cells could be found.. and. was. though. femur. same. at its later. of Experiments. endosteum. calcium,. be something. Experiment. of the. to undergo. of complete. - bination. is produced.. bone. calcium,. from. bone still has bone cells stained. the. must. 3, viz., bone dust transplantation.. no more stained. bone. cells trans. similar.. piece. this transplant,. the. percentage. bone. as well as endosteum. therefore, ment. that. tissue. it is supposed. periosteum. positive. the. and acts to the germ. artificial. The. it is removed. muscle. thus. with. artificial. are. soluble. tissue,. of the periosteum. positive. positions. becomes. connective. obtained. bone. host. that. and. this result. 7 and 9 showed. periosetum. Albee20 pointed. as well that. the. graft on bone transplantation should be autogeneous, consisting of all four bone layers; namely periosteum, endosteum, complete thickness of cortex and marrow. obvious. that. Considering Experiment. result, namely, which. naturally. 7 is to be certainly. the transplant consists. mount of bone part. 31. from the results. obtained. of periosteum,. The positive. of above experiments, accomplished. by strong currettment subperiosteal. percentage. layer. it is also. a satisfactory from the femur and. on this experiment. some. a. is supe.

(32) - 454rior. to that. bone. dust.. of Experiment These. things. is an auto-transplant. and. ctionless. osteoblasts. production that. regeneration so-called. dition. is existed.. The. fovourable dust. much,. mentioned.. However,. with. benefit,. disturbs. when the. adopted. bone. to the. dust. This. 8;. namely,. bone. substitutes. in the. for the. to help. transplant be. similar. bone. dust. ex in ad. furnishes. When. the. as. a. bone. hitherto. the removed former. dust. must. regeneration with. on. bone. layer. one.. coexists. regenerative. the. transplant. as previous. part. moreover,. advantage. in the the. the. fun. to bone. experiment. result. when. for the. the bone. beneficial, a great. be found. positive. bone. related. no subperiosteal. layer.. new. way. 4 is that. bone could. is superfluous. it surely. is insufficient,. new. obtained. to form. are. Experiment. in Experiment. one. environment. is too. steum. dust. previous. cells are not senile. is more. that. with. to the periosteum. are in some. process. and also. Goto15. bone. to the. on. No. bone. part. this. osteogenetic. extended.. periment.. bone. of periosteum. accompanies. that. which. assists. non-active. subperiosteal. furthermore. cells this. part. view. a disadvantage. is a homo-transplant,. transplantation. the bone. active. layer. 7 and. the. if Bart's30. but. subperiosteal. Experiment. of. that. is acknowledged,. the. the. 4, viz.,. and. perio will. be,. process.. The Experiment 9 was subjected from the standpoint that the endos teum has the same function with the periosteum. It is practically impos sible to remove endosteum alone as it is illustrated at Experiment 9. The Experiment 9 consists of endosteum, subendosteal layer, some amount of bone part and stillmore the bone marrow. The regenerative activity on this. experiment. written. in the. similar. tissue. was. text-book with. ment. 7, namely,. some. amount. marrow. the the. of bone. the bone marrow the differences. exceedingly that. the. periosetum. part. and. has. is consented,. assumed. must. a great. the. which same. result. is hitherto function. of the. subperiosteal. result. be considered. the Experiments. fact. the. of periosteum, an. exists or not, resulted. If the. endosteum. transplantation. is excluded. between. beneficial.. 7 and difference. Experi layer. of Experiment. and. 9 where. to be the same. 9 as to whether. and. Only. the bone. on bone regeneration. of. the two. In fact, the bone marrow cannot be excluded from the Experi ment 9 and therefore, This. test consists. for comparison,. of Experiment. the. Experiment. 8 plus bone. marrow;. 10 was performed. viz., periosteum, 32.

(33) -456-. subperiosteal result. layer, some. revealed. 10 are almost. amount. of bone part. that the proliferative the same.. bone dust or artificial or the artificial. bone calcium. bone dust in Experiment. results. on Experiment. are out of question.. on this occasion,. 8.. and the. activity of both the Experiments. The positive. bone calcium. and bone marrow,. Moreover,. 9 plus either The. bone dust. is also superfluous. it is obvious that. 10 plus bone dust can form new bone,. 9 and. having. as the. the Experiment. considered. from the above. results.. Judging from above experiments, it must be borne in mind that the existence of bone marrow plays an important role to the bone formation and consequently, it must dissolve the question how the bone marrow it self will be able to proliferate new bone or not and the Experiment. 11. reveals this question. Some literature added.. on the transplantation. Saltykow34 proved. for the first. of bone marrow. will be here. time, that the bone marrow. trans. planted in the subcutaneous tissue of an animal mostly dies, however, what was retained its vitality can regenerate marrow cells and from the endo. steum coexists with the transplanted marrow, new bone is formed.. Fran. genheiin35 also obtained a positive result on homo-transplantation of bone marrow. Stillmore, Miyauchi36 investigated that new bone was formed from the endosteum autotransplanted animal.. with bone marrow in the liver of the same. Tongu16 removed bone marrow and made an emulsion which he. injected subcutaneously or muscularly. 31. days.. to. 129. was. In days.. very. other He. much. regenerative. ments,. partly. sues. part. of it. plantation days 33. to. that. and. bone. generally of bone. 4 months. the. on. the. activity. bone. in the. it.. of the. is due. crushing. of the. The. necrosis.. Ueda37. in the. subcutane. and. to the. bone. to the. loss ele. of new. surrounding entire,. especially. observed. or in the. regeneration;. marrow. as a matrix. of the. marrow. from 8. of osteoblastic. marrow. surface. partly. was transplanted. marrow as. in toto. This. marrow. arround. underwent. was transplanted. of the. volume. increase. Where formed. former.. account. insufficient to the. marrow osteogenetic. in the. power. for absorption. of new. found. to the. formation,. a ring. the bone. decreased. of the. bone. cases,. The experiment lasted for 14 to. an. tis. there the. was. central. auto-trans. liver of rabbit, he, viewed , that. for 3 the.

(34) - 456osteoblasts seous. in the. be drawn bone. from. the. ists. hand,. this. contrary, the. with. the. uchi's36. marrow. bone. marrow. form. an os. attention. must. and. very. doubtful. From the. bone. ration. tuted. also rently. is some. quite fact. similar. of many the. itself. bone. the. increases. 10,. 11, no. occurs. marrow,. how. with. all. the. experiment. 9 it may. and. nutrient. form. to the. added. that. per. The. transplantation. be furthermore. it is or. bone. of bone. failed to form. investigated. but. it. coagulum for their Yokoi17. of fibrin. marrow. that. is substi. cases.. cent. not.. regene. mentioned,. He. must. of discus. bone,. but the blood. above. calcium. marrow. coagulum. in any. bone. of, at. considered. new. be found. or artificial. trans. removed be. blood. one. of Miya. described,. been. to. transplants. in. alone nor. is point. could. view.. dust. has. if the. fact. real bone. hitherto,. it will be resulted,. particurality. proliferation.. This. activity. is that. bone,. calcium. endosteum the. the. Ueda27 and the others'. one.. and. that. of cooperation. mean. marrow. coex. the endosteum. from. results,. for a stimulant. same. of bone this. bone. entirely. next. no. to above. part. the. a. the latter,. of normal. The. existed.. positive. real. has. associated. pointed. endosteum. and. bone. thus. from. not. of the brings. considered. as a product. does. of the bone.. new bone. action.. neither. bounda. this. the. the. be selected. periphery. to those. was formed. and. at their. the. and. in the. experiment. of Experiments. however,. either. endosteum,. His. also. part. be considered. and. question. always. However,. must. similar. requirement,. the new bone. serviceable. for the. nutrient,. take. to form. are. for its. probably. marrow. The. marrow. be applied. whether. is excellently. have. having. in bone. failed. rabbit,. must. accordingly. amount,. liver. views. with. and. small. as to the. the. in contact. endosteum. former. on. periphery. is quite. as there. sion. the. Even. that. are. from. bone.. alone.. experiments. apart. marrow the. least,. purpose. formed. marrow. transplantation. bone,. bone. An. with the endosteum transplanting. newly. may. in the bone. At. showed. will. least,. marrow.. bone. the. experiment. planted. results.. In fact,. composition. marrow. different. condition. with. endosteum. calcium. bone. the. quite. on this. of a long. associates. result.. calcium. with. associate. real. marrow. portion. other. with. of the. bone. always. different. two. is in inseparable. so the. the. showed. on removal. central. bone. on. 11. marrow. and. On. which. tissue. Experiment. ry. endosteum. appa. combined new bone.. as that. of Ex 34.

(35) - 457-. periment 2 as far as the bone marrow is an urgent stimulant and also it must be considered that this transplant ought to have the possibility on bone regeneration more than that of Experiment 2. The positive result on the transplantation of periosteum with either of bone dust or articial bone cal cium (Experiments 4 and 5) was quite in a minority and therefore, on Ex periment. 13,. ments. 4 or. cluded. that. a stimulant, 5 and the. the. bone. to accomplish. bone. was. and. too. small. cannot. ed,. was. By appendix, row. is absolutely. tation. that. is great. at least served,. it is well. Periosteum new. was. added. it can. number. of Experiment. the. result. and. for bone. that. no. for bone formation, in such. a place. con. and nutrient 14. must. and. wait. 14 and. 15. further 15 show. formation.. periosteum. and. to Experi. be definitely. for a stimulant. benefit,. understand. endosteum. as well. thus. as far as the experiments. nor. it may. where. bone. say without. there. mar hesi. is absolutely. existed.. VIII. 1.. marrow. essential. tell. with. can be formed. nor. The. definitely. essential. bone. no periosteum. not. bone. beneficial,. regeneration.. However,. spleen. the. was. marrow. investigation. the. namely, result. Conclusions.. as endosteum. itself. has. entirely. no. activity. to form. bone.. 2. However the cooperation of periosteum (endosteum) and bone calcium, when it furnishes a favourable environment, can be accomplished bone regeneration. 3.. Bone. marrow. as a stimulant. has and. also. no. nutrient. activity. to form. for bone. new. regeneration. bone. but it plays. a role. .. 4. Periosteum (endosteum) is not always necessarily on bone formation.. 35.

(36) -458-. IX.. Bibliography.. 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18.. Duhamel, from "Human embryology" by Kiebel and Mall. Virchow, from "Autoplastic Surgery" by Davison and Smith. Gegenbauer, Ditto. Oilier, Ditto. Moscheowitz, John Hopkins' Hospital Bull. XXVII, 1916. Maeewen, The growth of bone. 1912. Gedden, Brit. Med. Assoc. Sept. 28, 1912. Paterson, Ditto. Wm. Wright, Ditto. Douglas Crowford, Ditto. Mayer, and Wehner, Arch. f. Klin. Chirur. CIII 2pl, 1912. Gallie and Robertson, Brit. Med. J. Oct. VIII, 1919. Minoura, Frankf. Ztschr. f. Path. XV 3 pl., 1914. Nakahara, Arch. f. Orthopedic. VII, 1909. Goto, Mittel. aus d. Med. Fakultat d. Kaiser. Unives tat Kiushu B. 1, 1914. Tongu, Mittl. aus d. Med. Fak. d. Kaiser. Universitat Tokyo XIV, 1915. Yokoi, Deut. Zeitschr. f. Chirur. CXIII, 1912. Tsunoda, Tokyo Ijishinshi. No. 1760.. 19. 20. 21. 22. 23.. Murphy, J. Am. Med.Assoc. April 6, 1921. Albee, J. Am. Med.Assoc. April 5, 1913. Graves, Brit. J. Surg. Vol. 1, No. 3, 1914. Fujimori, Nippon Geka Gakkai. No. 13. Tomita, VirchowArch. f. Path. Anat. 191, 1908.. 24. 25. 26. 27. 28. 29. 30. 31. 32. 33. 34. 35. 36. 37. . 38. 39. 40.. Ryerson, Canada Med. Assoc. J. III, 1913. Bancroft, Am. J. Med. Soc. CXLVII, 1914. Davison and Smith, Autoplastic Surgery. 1916. Ely, Annal Surg. 70, 1919. Hyde, Prc. Connecticut State Med. Soc. CXXVIII, 1920. Bonome, Virchow Arch. f. Path. Anat. 100, 1895. Bart, Am. J. Anat. 29, 1920. Carnot, from Chemical Pathology by T_??_unoda.p. 424. Asami and Dock, J. Exp. Med. Dec. 1, 1920. Koga, Kyoto Igakkai Zasshi. Bd. 9, No. 2, 85, 1917. Saltykow, Ziegl. Beitr. Bd. 45, 1909. Frangenheim, Arch. f. Klin. Chirur. No. 93, 1910. Miyauchi, Arch. f. Klin. Chirur. Bd. 106, 1913. Ueda, Chugai Iji Shinpo. No-891,1917 Nakayama, Nippon Geka Gakkai. Bd. 2, 1901. Harvey, J. Med. research. Oct. Vol. XVII (new series Vol. XII), 1907. Liek, Langenbeck Arch. f. Klin. Chirur. Bd. 80, 1906. Lewis and Stohr, A text book of Histology. Minot, Text book of Human Embryology. Kiebel and Mall, Human Embryology. Caningham, Text book of Anatomy. Dunham, Normal Histology. Stohr, Lehrbuch der Histologie. 36.

(37) -459-. X. Figure. 1.. No. 55 A.. Description. Transplantation. of Plates.. of periosteum. plus bone dust.. A quite large area of newly formed bone which possesses paration. fi eld. Figure 2.. bone marrow. is seen in this pre. Several groups of foreign body giant cells are also seen at the lower. No. 47 A.. Transplantation. of periosteum. part of the. plus bone dust.. Some foreign body giant cells in the fatty tissue are seen at the left side of the field.. A. new bone occupies the central part. Figure. 3. Dart. No.. There Figure. 4.. 56 A.. Figure. 5.. groups. 81 B. the. giant No.. of periosteum,. subperiosteal. layer,. some. amount. of bone. dust.. are two No.. part. Between body. Transplantation. and bone. of newly. formed. Transplantation. two. insels. of. bone. which. of periosteum,. newly. formed. possess. some. suhperiosteal. bone,. there. bone. layer,. are. seen. marrow some. several. are shown.. amount. of. groups. of. bone. foreign. cells. 19 B.. Transplantation. part and bone A ring formed. marrow. new bone. blood-corpuscles. is also. of endosteum,. tissue seen. is seen. in the. at one side. subendosteal. fatty. of the. layer,. tissue.. Some. some. bone. amount. marrow. of. rich. bone. in. red. ring.. Figure 6. No. 84 B. Transplantation of bone marrow(periphery). The appearanceis quite similar to that of previouscase. Figure 7. No. 60 A. Transplantationof periosteum,subperiosteallayer, some amount of bone part and bone marrow. The field is occupiedwith a quite large area of newly formedbone. No bone marrow nor giant cells can be seen in this preparate. Figure. 8.. No. 74 A.. Transplantation. of periosteum,. subperiosteal. layer,. part and bone marrow. A small rounded area of newly formed bone is seen in the fibrous great deal of giant cells-groups Figure. 9. part, New. No.. 53 A.. bone bone. Figure. 10.. formed. is scarecelv 11.. No.. Figure. 37. 12.. Transplantation bone. seen. calcium. new group. No. 19 A.. suhperiosteal. and some. of periosteum,. occupies. at the lower. 68 A.. A semicircled This. of periosteum,. layer,. foreign. body. Besides. this, a. bed. some. amount. giant. cells. are. of bone. located. of the field.. No. 62 A.. A newly. Figure. Transplantation. Dart. tissue.. occupies nearly all the transplanted. dust and bone marrow. groups are exhibited in the field. at the lower. some amount of bone. the central. of periosteum,. is situated. is nearly. bone. dust and. of the field. and. artificial. bone. bone. marrow.. some. degenerated. marrow. Dart of the field.. Transplantation bone. area. in the. area. which. consists. calcium. and bone. of artificial. bone. marrow. calcium.. homogeneous.. layer,. some amount of bone. part, bone marrow and artificial bone calcium. A quite large area of newly formed bone is seen and no artificial found.. Transplantation. of endosteum,. subendosteal. bone calcium can be.

(38) Fig.. 1.. Fig.. 2.. Fig.. 3.. Fig.. 4.. Fig.. 5.. Fig.. 6..

(39) Fig.. 7.. Fig.. 8.. Fig.. 9.. Fig.. 10.. Fig.. 11.. Fig.. 12..

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岡 山醫學 會雜誌 第 四百 十 二號 大 正 十 三 年 五 月 三 十 一 日發 行

岡山醫學會雜誌第四百十二號大正十三年五月三十一日發行