This study has been performed to evaluate the relationship between the remained mineral components in a decalcified bone matrix and an ectopic bone formation efficiency. The freezed rat diaphyseal cortical bones measuring 0.5cm in length were demineralized in heated 0.6N HCl at $60^{\circ}C$ for 5, 10, 15, 20, 25, 30, 35, 40 minutes, respectively, using a controlled heat ultrasonic cleaner. Each 1cc of decalcifying solution taken during decalcification procedure was used to calculate calcium content using calcium dignostics kit under 600nm of spectrophotomer. After decalcification, each specimen was also weighed. Then each prepared specimen was implanted into the dorsal pouch of 24 Sprague-Dawley rats divided into 8 groups by time course. The implants were harvested at 1, 2, and 3 weeks and prepared for routine H-E stain specimens to evaluate osteogenic activity. The results are as follows: 1. There was statistical significant difference in change of calcium concentration up to demineralization of 30 minutes and each allogenic bones decalcifed up to 20 minutes revealed 99.65% of decalcification in average. 2. There was statistical significant difference in change of weight in demineralized allogenic bone up to 20 minutes treatment but, no significant change was noted after that time. 3. The histologic analysis revealed active ectopic bone formation in the implanted allografts demineralized for 20, 25, 30 minutes, respectively. However, the other groups of allografts showed relatively poor osteoinductive activity. These findings suggest that complete decalcification with a minimized degeneration of collagen matrix is necessary to induce maximal osteogenesis by decalcified bone allograft.
DLA class I complement-dependent cytotoxicity (CDC) cross-match method was established to control hyperacute rejections in organ transplantation. The aim of the present study is to investigate if DLA class 1 CDC corssmatch method is effective to prevent hyperacute rejections in canine renal allografts. Seven mongrel dogs of similar age and weight were used. Erythrocyte crossmatch was first performed and only the negatives were used. Among the same blood types, CDC cross-match was performed. Anti-dog serum, Hank's balanced salt solution(HBSS), and tile self-serum was used as a positive-, a negative-, and all auto-control respectively. After the reaction with class I complement, it was stained with eosin and scored by international cytotoxicity scoring system under inverted phase microscope. According to these results, kidneys oi CDC negatives among same blood types were cross-transplanted to observe the incidence of hyperacute rejections. One of four 1.2 B blood type dogs had autoantibodies. here were negative CDC results among each blood type, and also there were negative results between different blood types. Two pairs with the same blood types and negative CDC results underwent allo-transplantation each other. There were no hyperacute rejections. DLA cross-match method studied in this experiment for canine renal allograft can be effective to prevent hyperacute rejections. it may be applicable for the future studies of histocompatibility testing in canine renal allografts.
Purpose : Preliminary report of the technique and trial of double bundle PCL reconstruction using Achilles allograft. Materials and Methods : From May 1999 to July 2000, 8 cases of PCL insufficient patients were treated with Achilles allograft reconstruction using the double bundle and double femoral tunnel technique. The tibial tunnel was prepared anteromedially. All other combined injuries within the knees were treated accordingly. Minimal follow-up period was 1 year. The results was assessed from the point of function and stability using Lysholm knee score and KT-2000 arthrometer. Results : Up to present follow up. 8 patients showed good sign of recovery with no instability (translation less than 2 mm) except olio that has been grafted-ligament rupture. In addition, none showed any sign of infection nor ROM limitation. Two complications were seen, which one had grafted-bone fracture and the other grafted-ligament rupture. The former occurred during operation and the latter occurred due to improper protection. Conclusion : Presently the follow up period is too short to draw any conclusive opinion but it is essential to select healthy and well sterilized allografts fur successful outcome. Double femoral tunnel technique seems to be more physiologic in PCL reconstruction. With these prerequisites, it seems to be a good alternative to use Achilles allografts fur the reconstruction of PCL. However, a longer follow-up is needed.
Woo, Joon-Bum;Son, Dong-Wuk;Lee, Su-Hun;Lee, Jun-Seok;Lee, Sang Weon;Song, Geun Sung
Journal of Korean Neurosurgical Society
/
v.62
no.4
/
pp.450-457
/
2019
Objective : Anterior cervical discectomy and fusion (ACDF) is commonly used surgical procedure for cervical degenerative disease. Among the various intervertebral spacers, the use of allografts is increasing due to its advantages such as no harvest site complications and low rate of subsidence. Although subsidence is a rare complication, graft collapse is often observed in the follow-up period. Graft collapse is defined as a significant graft height loss without subsidence, which can lead to clinical deterioration due to foraminal re-stenosis or segmental kyphosis. However, studies about the collapse of allografts are very limited. In this study, we evaluated risk factors associated with graft collapse. Methods : We retrospectively reviewed 33 patients who underwent two level ACDF with anterior plating using allogenous bone graft from January 2013 to June 2017. Various factors related to cervical sagittal alignment were measured preoperatively (PRE), postoperatively (POST), and last follow-up. The collapse was defined as the ratio of decrement from POST disc height to follow-up disc height. We also defined significant collapses as disc heights that were decreased by 30% or more after surgery. The intraoperative distraction was defined as the ratio of increment from PRE disc height to POST disc height. Results : The subsidence rate was 4.5% and graft collapse rate was 28.8%. The pseudarthrosis rate was 16.7% and there was no association between pseudarthrosis and graft collapse. Among the collapse-related risk factors, pre-operative segmental angle (p=0.047) and intra-operative distraction (p=0.003) were significantly related to allograft collapse. The cut-off value of intraoperative distraction ${\geq}37.3%$ was significantly associated with collapse (p=0.009; odds ratio, 4.622; 95% confidence interval, 1.470-14.531). The average time of events were as follows: collapse, $5.8{\pm}5.7months$; subsidence, $0.99{\pm}0.50months$; and instrument failure, $9.13{\pm}0.50months$. Conclusion : We experienced a higher frequency rate of collapse than subsidence in ACDF using an allograft. Of the various preoperative factors, intra-operative distraction was the most predictable factor of the allograft collapse. This was especially true when the intraoperative distraction was more than 37%, in which case the occurrence of graft collapse increased 4.6 times. We also found that instrument failure occurs only after the allograft collapse.
Purpose: The aim of the present study was to evaluate the effectiveness of using a mineralized bone cortical allograft (MBCA), with or without a resorbable collagenous membrane derived from bovine pericardium, on alveolar bone remodeling after immediate implant placement in a dog model. Methods: Six mongrel dogs were included. The test and control sites were randomly selected. Four biradicular premolars were extracted from the mandible. In control sites, implants without an allograft or membrane were placed immediately in the fresh extraction sockets. In the test sites, an MBCA was placed to fill the gap between the bone socket wall and implant, with or without a resorbable collagenous membrane. Specimens were collected after 1 and 3 months. The amount of residual particles and new bone quality were evaluated by histomorphometry. Results: Few residual graft particles were observed to be closely embedded in the new bone without any contact with the implant surface. The allograft combined with a resorbable collagen membrane limited the resorption of the buccal wall in height and width. The histological quality of the new bone was equivalent to that of the original bone. The MBCA improved the quality of new bone formation, with few residual particles observed at 3 months. Conclusions: The preliminary results of this animal study indicate a real benefit in obtaining new bone as well as in enhancing osseointegration due to the high resorbability of cortical allograft particles, in comparison to the results of xenografts or other biomaterials (mineralized or demineralized cancellous allografts) that have been presented in the literature. Furthermore, the use of an MBCA combined with a collagen membrane in extraction and immediate implant placement limited the extent of post-extraction resorption.
To determine the sterility of the prepared allogeneic bone of the human, culture of the allografts prior to implantation was performed on fresh-frozen and freeze-dried bone. Before the use of allografts to the patients, it must be confirmed about the sterility, cellular cytotoxicity, immune reaction, and osteoinductive potential as a biomaterials. Oral and maxillofacial surgeons demand for allograft bone will be increased in the future. Wonkwang Bone Bank attempted to meet this demand, has performed series of experimental study on the allograft bone of the Koreans to evaluate the physical and chemical suitability of the bone since the surgeons applications will have broadened from benign cystic lesions to fracture malunions and non-unions, large segmental defects, and whole-bone allorgrafts after tumor surgery. The results obtained were as follows: 1. Freeze-drying(FD) only shoed some bactericidal effects of the normal and osteo bone but in cases of performing EO gas sterilization, the FD effects was not clear. 2. The fact that FD has little effect than the EO gas sterilization on normal bone postulated that the presence of microbiota may be due to an operation and bone processing procedure. 3. FD and EO gas sterilization had a remarkable effect on the osteo bone. 4. The sterilization effect were EO gas, Freeze-drying, Fresh-Frozen with descending order. But all sterilization method were not complete to preserve and implant allograft bone. We are now performing further continuous study on the radiation and chemical sterilization procedure to make safe and complete allograft bone.
Implantation of allografts has increased widely with not only the availability of many allogenic bone but also allogenic soft tissues. The aim of tissue banking is to provide surgeons with safe tissues compatible with their intended clinical application. The incidence of tissue transplant-transmitted infection is unknown and can only be inferred from prospective studies. The possibility of donor-to-recipient disease transmission through soft tissue transplantation can be considered by reviewing the risk associated with other transplanted hard tissues. Viral, bacterial, and fungal infections have been transmitted via transplantation of soft tissue allografts such as skin, cornea, dura, pericardium. fascia lata, and heart valves. Corneas have transmitted rabies, Creutzfeldt-Jakob disease (CJD), hepatitis B (HBV), cytomegalovirus (CMV), herpes simplex virus (HSV), bacteria, and fungi. Heart valves have been implicated in transmitting tuberculosis, hepatitis B. HIV-1 and CMV. CJD has been transmitted by dura and pericardium transplants. Skin has transmitted CMV, bacteria, and fungi. Cadaveric skin, pericardium, dura, and fascia lata have been used in dental patients with intra-oral soft tissue injuries and GBR. This study is review of the considering transmission of infectious disease in allogenic soft tissues and guidelines of reducing the risk. Prior to use, many tissues are exposed to antibiotics, disinfectants, and sterilants, which further reduce or remove the risk of transmitted disease. Because some soft tissue grafts cannot be subjected to sterilization steps, the risk of infectious disease transmission remains and thorough donor screening and testing is especially important.
Bone allograft had been increased the need because autogenous bone graft is associated with donor site morbidity and is restricted in quantity. The bone allograft implants have to prepare properly for increasing osteoinductive ability and decreasing immune responses before providing to graft. The purpose of this study was to investigate the efficacy on new bone formation in bone allografts by treatment of implants. Cortical bone allografts were transplanted to experimental defects on midshaft of diaphyseal fibulae in 15 rabbits, which were divided to 3 experimental groups according to the preparation methods-freezing, freeze-drying, defat-freezing. The grafted regions of fibulae of all groups had been radiographed biweekly for 16 weeks to observe new bone formation and union between implant and recipient bone. Bone Alkaline Phosphatase (BALP) in all groups was evaluated biweekly till the end of the experiment to determine osteoblast activities. Unions between implant and recipient bone were observed at 30% (3 of 10 cases) of freezing, 50% (5 of 10 cases) of freeze-drying and 80% (8 of 10 cases) of defat-freezing. BALP was increased over 100% from before graft at 2 weeks of graft procedures in all union cases of freezing and defat-freezing group, then gradually decreased till 16th week. In non-union cases, there is no significant variation in BALP value. Defat-freezing method for allograft implants might be more effective for osteoinductive efficacy of implants than freezing and freeze-drying method.
Eun, Seok Chan;Kim, Byung Jun;Kim, Jin Hee;Heo, Chan Yeong;Baek, Rong Min;Chang, Hak;Minn, Kyung Won
Archives of Plastic Surgery
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v.35
no.4
/
pp.367-372
/
2008
Purpose: Prevention of acute rejection in skin allografts without continuous immunosuppression lacks reports in worldwide literature. Needs for chronic immunosuppression preclude the use of tissue allograft as a routine surgical reconstructive option. Recently dendritic cells(DC) gained considerable attention as antigen presenting cells that are also capable of immunologic tolerance induction. This study assesses the effects of alloantigen-pulsed dendritic cells in induction of survival increase in a rat skin allograft model. Methods: Recipient-derived dendritic cells were harvested from rat whole blood and cultured with GM-CSF(200 ng/mL) and IL-4(8 ng/mL) for 2 weeks. Then donor-specific alloantigen pulsed dendritic cells were reinjected into tail vein before skin graft. The rat dorsal skin allografts were transplanted in 5 subgroups. Groups: I) untreated, II) anti-lymphocyte serum(ALS, 0.5 mL), III) FK-506(2 mg/kg), IV) DCp, VI) DCp and FK-506. Graft appearance challenges were assessed postoperatively. Results: The group V(DC and FK-506 treated) showed longest graft survival rate(23.5 days) than other groups; untreated(5.8 days), ALS(7.2 days), FK-506 (17.5 days), DCp(12.2 days). Conclusion: Donor antigen pulsed host dendritic cell combined with short-term immunosuppression prolong skin allograft survival and has potential therapeutic application for induction of donor antigen specific tolerance.
Kanatani, Takako;Fujioka, H.;Lanzetta, M.;Kurosaka, M.;Matsumoto, T.;Bishop, G.A.
Archives of Reconstructive Microsurgery
/
v.18
no.1
/
pp.9-15
/
2009
Whether a seven days course of anti-${\alpha}{\beta}$-T cell receptor-antibody (${\alpha}{\beta}$-TCRmAb) combined with FK506 therapy promotes survival of limb allografts in fully MHC-mismatched combination (Brown Norway $\rightarrow$ Lewis) was examined. Eight animals received 250 ${\mu}g$/kg/day of ${\alpha}{\beta}$-TCRmAb for 7 days and 2 mg/kg/day of FK506 postoperatively (Combination therapy group). Eight animals had FK506 only (Mono-therapy group) and five animals did not have treatment (Control group). Clinical signs of early rejection with edema or erythema in the skin occurred at an average of 8.6${\pm}$1.5 days postoperatively in Control group and 59.0${\pm}$8.3 days in Mono-therapy group, both of which proceeded to irreversible rejection with necrosis of the epidermis and finally mummification. In Combination therapy group, all animals showed evidence of early rejection at an average of 56.8${\pm}$12.6 days postoperatively, however, in 4 of 8 limbs, early rejection resolved without any treatment and limbs survived >1 year. At 9 months postoperatively, donor skin grafts were accepted and third-party skin grafts were rejected by all four survivors, demonstrating donor-specific tolerance. Little or no detectable chimerism was observed in any of the 4 surviving animals at one-year postoperatively. Combination therapy of ${\alpha}{\beta}$-TCRmAb and FK506 resulted in long-term survival in fully MHC-mismatched limb transplants.
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