PHYSICOCHEMICAL CHARACTERIZATION OF LYOPHILIZED BOVINE BONE GRAFTS.

UNLABELLED: To evaluate the physicochemical characteristics of lyophilized bovine grafts manufactured on a semi-industrial scale (Orthogen; Baumer S/A*) in accordance with a protocol previously developed by the authors. METHODS: The lyophilized bovine bone grafts were characterized by means of scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), X-ray diffractometry (XRD), thermogravimetric (TG) analysis, differential exploratory scanning calorimetry (DSC) and Fourier-transform infrared (FT-IR) spectroscopy. RESULTS: Ca was the main component (60%) found in the samples, followed by P (28%) and O (5%). The mean (sd) pore size was 316 µm (146.7), ranging from 91.2 to 497.8 µm, and 333.5 µm (304.8), ranging from 87.2 to 963.9 µm, at 50x and 150x magnification, respectively. The hydroxyapatite peaks were at 26°C and 32°C, and mass losses were observed between 250°C and 640°C, corresponding to organic material and water. Two temperature transitions (45.67°C and 91.89°C) showed denaturation of type 1 collagen and dehydration of hydroxyapatite. CONCLUSION: The physicochemical assessment of lyophilized bovine bone grafts in accordance with the protocol developed at semi-industrial scale confirmed that this product presents excellent biocompatibility, with characteristics similar to natural bone.

Caracterização físico-química do enxerto de osso bovino liofilizado / Physicochemical characterization of lyophilized bovine bone grafts

OBJETIVO: Avaliar as características físico-químicas do enxerto bovino liofilizado manufaturado em escala semi-industrial (OrthoGen, Baumer S/A*) de acordo com protocolo previamente desenvolvido pelos autores. MÉTODOS: A caracterização do enxerto de osso bovino liofilizado foi feita por meio de microscopia eletrônica de varredura (MEV), energy dispersive spectroscopy (EDS), difratometria de raios-X, análise por termogravimetria, análise de calorimetria exploratória diferencial (DSC) e espectroscopia por infravermelho Fourier-transform (FT-IR). RESULTADOS: Ca foi o principal componente (60 por cento) encontrado nas amostras, seguido por P (28 por cento) e O (5 por cento). O tamanho médio (dp) dos poros foi 316µm (146,7), variando de 91,2 a 497,8µm, e 333,5µm (304,8), variando de 87,2 a 963,9µm com 50x e 150x magnificação, respectivamente. Picos de hidroxiapatita foram a 26ºC e 32ºC, e perda de massa foi observada entre 250ºC e 640ºC, correspondendo material orgânico e água. Duas transições de temperatura (45,67°C e 91,89°C) mostraram desnaturação de colágeno tipo I e desidratação da hidroxiapatita. CONCLUSÃO: A avaliação físico-química do enxerto de osso bovino liofilizado, de acordo com o protocolo desenvolvido em escala semi-industrial, confirma que este produto apresenta excelente biocompatibilidade, com características semelhantes ao osso in natura.

OBJECTIVE: To evaluate the physicochemical characteristics of lyophilized bovine grafts manufactured on a semi-industrial scale (Orthogen; Baumer S/A*) in accordance with a protocol previously developed by the authors. METHODS: The lyophilized bovine bone grafts were characterized by means of scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), X-ray diffractometry (XRD), thermogravimetric (TG) analysis, differential exploratory scanning calorimetry (DSC) and Fourier-transform infrared (FT-IR) spectroscopy. RESULTS: Ca was the main component (60 percent) found in the samples, followed by P (28 percent) and O (5 percent). The mean (sd) pore size was 316 µm (146.7), ranging from 91.2 to 497.8 µm, and 333.5 µm (304.8), ranging from 87.2 to 963.9 µm, at 50x and 150x magnification, respectively. The hydroxyapatite peaks were at 26°C and 32°C, and mass losses were observed between 250°C and 640°C, corresponding to organic material and water. Two temperature transitions (45.67°C and 91.89°C) showed denaturation of type 1 collagen and dehydration of hydroxyapatite. CONCLUSION: The physicochemical assessment of lyophilized bovine bone grafts in accordance with the protocol developed at semi-industrial scale confirmed that this product presents excellent biocompatibility, with characteristics similar to natural bone.

Femoral and acetabular revision using impacted nondemineralized freeze-dried bone allografts.

BACKGROUND: Favorable results have been obtained by the use of deep-frozen bone allografts in total hip arthroplasty. However, owing to the shortage of deep-frozen allografts and the risk of infectious disease, other materials have been studied, such as sterile nondemineralized freeze-dried allografts. The aim of this study was to describe midterm clinical outcomes and radiographic bone incorporation of human freeze-dried bone grafts in 42 revision total hip arthroplasty procedures using cancellous impacted bone grafting. METHODS: This report presented clinical and radiographic evidence of allograft incorporation in 42 hip reconstructions performed between 1996 and 2002. The patient group included 13 (31%) men and 29 (69%) women with mean +/- SD age of 63 +/- 14 years (range 28-80 years). Mean follow-up was 82 months (range 63-127) months. Clinical analysis was based on the D'Aubigné-Postel score. Radiographic incorporation was defined according to specific criteria. RESULTS: The D'Aubigné and Postel criteria showed adequate outcome in 38 (90%) of the patients. The radiographic evaluation revealed that allograft remodeling and incorporation were found in 39 (93.0%) and 36 (86.5%) of acetabular and femoral cases, respectively. The overall graft survival rate at an average follow-up of 8 years (range 5-10 years) was 90%. CONCLUSIONS: Bone grafts obtained by the lyophilization process developed and carried out in our tissue bank provide suitable grafts for revision total hip arthroplasty. Clinical and radiographic midterm results were excellent, indicating that nondemineralized freeze-dried bone allografts are suitable for replacing deep-frozen grafts.

In vitro and in vivo evaluation of lyophilized bovine bone biocompatibility.

INTRODUCTION: The use of bone grafts in orthopedic, maxillofacial and dental surgery has been growing. Nevertheless, both fresh autografts and frozen allografts have limitations, and therefore, alternative synthetic or natural biomaterials, such as processed and lyophilized bovine bone graft have been developed. OBJECTIVE: To evaluate in vitro and in vivo biocompatibility of lyophilized bovine bone manufactured in a semi-industrial scale, according to a modifical protocol developed by the authors. METHODS: Samples of bovine cancellous bone were processed according to a protocol developed by Kakiuchi et al., and modified to process samples of bovine cancellous bone. The following trials were performed: in vitro cytotoxicity, in vivo acute systemic toxicity, in vivo oral irritation potential, in vitro pyrogenic reaction, and bioburden. RESULTS: The in vitro evaluation of lyophilized bovine cancellous bone revealed an absence of cytotoxicity in 100% of the samples. Regarding in vivo evaluation of acute systemic toxicity, neither macroscopic abnormalities nor deaths were noted in the animals. Pyrogenicity was not greater than 0.125 UE/ml in any of the samples. The bioburden revealed negative results for microbial growth before sterilization. Regarding the oral irritation potential, in vivo evaluation at 24 and 72 hours showed that the animals had no edema or erythema on the oral mucosa. CONCLUSION: The protocol changes established by the authors to prepare lyophilized bovine cancellous bone at a semi-industrial scale is reproducible and yielded a product with excellent biocompatibility.

In vitro and in vivo evaluation of lyophilized bovine bone biocompatibility

INTRODUCTION: The use of bone grafts in orthopedic, maxillofacial and dental surgery has been growing. Nevertheless, both fresh autografts and frozen allografts have limitations, and therefore, alternative synthetic or natural biomaterials, such as processed and lyophilized bovine bone graft have been developed. OBJECTIVE: To evaluate in vitro and in vivo biocompatibility of lyophilized bovine bone manufactured in a semi-industrial scale, according to a modifical protocol developed by the authors. METHODS: Samples of bovine cancellous bone were processed according to a protocol developed by Kakiuchi et al., and modified to process samples of bovine cancellous bone. The following trials were performed: in vitro cytotoxicity, in vivo acute systemic toxicity, in vivo oral irritation potential, in vitro pyrogenic reaction, and bioburden. RESULTS: The in vitro evaluation of lyophilized bovine cancellous bone revealed an absence of cytotoxicity in 100 percent of the samples. Regarding in vivo evaluation of acute systemic toxicity, neither macroscopic abnormalities nor deaths were noted in the animals. Pyrogenicity was not greater than 0.125 UE/ml in any of the samples. The bioburden revealed negative results for microbial growth before sterilization. Regarding the oral irritation potential, in vivo evaluation at 24 and 72 hours showed that the animals had no edema or erythema on the oral mucosa. CONCLUSION: The protocol changes established by the authors to prepare lyophilized bovine cancellous bone at a semi-industrial scale is reproducible and yielded a product with excellent biocompatibility.