Human placenta-derived matrix with cancellous autograft and demineralized bone matrix for large segmental long-bone defects in two dogs with septic nonunion.

OBJECTIVE: To report the successful treatment of septic nonunion in two dogs with large segmental defects secondary to long-bone fractures by using a novel human placenta-derived matrix (hPM) as adjunct to fixation. ANIMALS: One 3-kg 9-year-old neutered male Yorkshire terrier with a distal antebrachial fracture and one 6-kg 4-year-old spayed female miniature pinscher with a distal humeral fracture. STUDY DESIGN: Short case series. METHODS: Both dogs presented for septic nonunion after internal fixation of Gustilo type II open diaphyseal fractures from dog bite injuries. During revision, debridement of nonviable bone resulted in segmental defects of 32% and 20% of the bone length for the antebrachial and humeral fractures, respectively. The antebrachial fracture was stabilized with a circular external fixator, and the humeral fracture was stabilized with biaxial bone plating. The fracture sites were not collapsed, and full length was maintained with the fixation. Autogenous cancellous bone graft and canine demineralized bone allograft were packed into the defects, and hPM was injected into the graft sites after closure. RESULTS: Radiographic union was documented at 8 weeks and 6 weeks for the antebrachial and humeral fractures, respectively. Both dogs became fully weight bearing on the affected limbs and returned to full activity. CONCLUSION: Augmenting fixation with grafts and hPM led to a relatively rapid union in both dogs reported here.

Osteogenic potential of mesenchymal cells derived from canine umbilical cord matrix co-cultured with platelet-rich plasma and demineralized bone matrix.

Canine mesenchymal cells (MSCs) derived from Wharton's jelly were co-cultured, then supplemented or not supplemented with platelet rich plasma (PRP) and demineralized bone matrix (DBM) to verify osteogenic differentiation. Osteoblastic differentiation followed by mineralized bone matrix production was found to be significantly higher (p < 0.05) when MSCs were associated with PRP/DBM in culture after 14-21-days of induction. Osteopontin and osteocalcin gene expression were significantly superior (p < 0.05) under the same culture conditions after 21 days of observation. In conclusion, addition of PRP to DBM co-cultured with MSCs successfully induced osteogenesis in vitro.

Evaluation of an Osseous Allograft Membrane for Guided Tissue Regeneration in the Dog.

Clinical application of a demineralized freeze-dried cortical bone membrane allograft (DFBMA) for treatment of intra(infra)bony periodontal pockets in dogs was evaluated. The mean pre-treatment periodontal probing depth equaled 7.2-mm. Post-treatment probing depths in all 11 cases were normal, with a mean periodontal probing gain of 5.4-mm. Guided tissue regeneration using a commercially available veterinary canine DFBMA and canine demineralized freeze-dried bone allograft (DFDBA) resulted in clinically significant periodontal attachment gains. The gain of new periodontal tissue attachment was statistically significant (P < 0.0001). The commercially available veterinary allograft products predictably increased new periodontal attachment without any identified membrane sequelae in these 11 cases.

Osteogenic potential of mesenchymal cells derived from canine umbilical cord matrix co-cultured with platelet-rich plasma and demineralized bone matrix

Canine mesenchymal cells (MSCs) derived from Wharton's jelly were co-cultured, then supplemented or not supplemented with platelet rich plasma (PRP) and demineralized bone matrix (DBM) to verify osteogenic differentiation. Osteoblastic differentiation followed by mineralized bone matrix production was found to be significantly higher (p < 0.05) when MSCs were associated with PRP/DBM in culture after 14-21-days of induction. Osteopontin and osteocalcin gene expression were significantly superior (p < 0.05) under the same culture conditions after 21 days of observation. In conclusion, addition of PRP to DBM co-cultured with MSCs successfully induced osteogenesis in vitro.

Qualitative and quantitative evaluation of avian demineralized bone matrix in heterotopic beds.

OBJECTIVE: To evaluate the osteogenic potential of avian demineralized bone matrix (DBM) in the context of implant geometry. STUDY DESIGN: Experimental. ANIMALS: Rock pigeons (n = 24). METHODS: Tubular and chipped forms of DBM were prepared by acid demineralization of long bones from healthy allogeneic donors and implanted bilaterally into the pectoral region of 24 pigeons. After euthanasia at 1, 4, 6, 8, 10, and 12 weeks, explants were evaluated histologically and compared by means of quantitative (bone area) and semi quantitative measures (scores). RESULTS: All explants had new bone at retrieval with the exception of tubular implants at the end of week 1. The most reactive part in both implants was the interior region between the periosteal and endosteal surfaces followed by the area at the implant-muscle interface. Quantitative measurements demonstrated a significantly (P = .012) greater percentage of new bone formation induced by tubular implants (80.28 ± 8.94) compared with chip implants (57.64 ± 3.12). There was minimal inflammation. CONCLUSIONS: Avian DBM initiates heterotopic bone formation in allogeneic recipients with low grades of immunogenicity. Implant geometry affects this phenomenon as osteoconduction appeared to augment the magnitude of the effects in larger tubular implants.

Potencial osteoindutor da matriz óssea homóloga desmineralizada de coelho / Osteoinductivity of demineralized homologous bone graft in rabbits

O trabalho objetivou avaliar o potencial osteoindutor da matriz óssea homóloga (MOD) desmineralizada em ácido clorídrico. Em doze coelhos adultos, foi realizada uma incisäo sagital mediana na pele abdominal, de aproximadamente 5 cm, e a divulsäo do tecido subcutâneo, permitindo que três fragmentos de MOD, com 1 cm de comprimento, fossem implantados em "bolsas" confeccionadas cirurgicamente no músculo reto abdominal. A MOD foi preparada a partir de rádios de coelhos sacrificados por superdosagem anestésica e estocada em etanol 70 por cento, em temperatura ambiente, por, no mínimo, 15 dias, até o momento do uso. Para permitir análise prévia às enxertias, fragmentos de MOD preservados em etanol 70 por cento e fixados em formol a 10 por cento tamponado foram submetidos à rotina histológica. Quatro coelhos foram sacrificados a cada duas, quatro e seis semanas de pós-operatório. A rotina histológica foi realizada, também, em dois dos enxertos implantados em cada animal, em cortes longitudinais e transversais, corados por H&E e no fragmento restante a deposiçäo de osso novo foi evidenciada pela fluorescência da tetraciclina administrada aos animais. Os resultados da pesquisa permitiram concluir que o ácido clorídrico 0,6 N promoveu desmineralizaçäo eficaz e manteve o potencial indutor da matriz óssea; a imersäo da matriz óssea desmineralizada em etanol 70 por cento propiciou a esterilizaçäo dos enxertos homólogos e näo inibiu o potencial osteoindutor; e também que, a MOD, devido a fatores indutores inerentes que atuam sobre as células dos tecidos adjacentes, promoveu quimiotaxia, mitose e diferenciaçäo celular.

Matriz óssea homóloga desmineralizada na preparaçäo de falhas ósseas segmentares produzidas no rádio de coelhos / Repair of segmental defects produced in the radii rabbit by homologous demineralized bone matrix

No presente trabalho, foram avaliados os potenciais osteoindutor e osteocondutor da matriz óssea homóloga desmineralizada (MOD) em ácido clorídrico 0,6 N e conservada em etanol 70 por cento e sua utilizaçäo no preenchimento de falhas osteoperiosteais induzidas em rádio de coelhos. O rádio operado de todos os animais foi radiografado imediatamente após a cirurgia e na 3ª, 6ª, 9ª e 12ª semanas após. No grupo tratado, sete animais foram sacrificados a cada três semanas, sendo realizadas tomografia computadorizada e avaliaçöes macro e microscópica e da fluorescência óssea à tetraciclina. Os animais do grupo controle foram sacrificados na 12ª semana de pós-operatório. Foi constatado que a matriz acelerou o processo de formaçäo óssea, devido a fatores indutores inerentes que atuaram sobre as células dos tecidos adjacentes e que os canais vasculares, as lacunas e o canal medular vazios da MOD, serviram de arcabouço para a penetraçäo celular, determinando osteoconduçäo.

Equine demineralized bone matrix: relationship between particle size and osteoinduction.

The osteoinductive capability of four particle sizes of equine demineralized bone matrix (DBM) was evaluated. Matrix particles were implanted in brachiocephalicus muscle pouches in six horses and were harvested 8 weeks later. Matrix particle sizes of 2.0 mm3 to 4.0 mm3 and 5.0 mm3 to 10.0 mm3 were associated with osteoinductive activity and minimal signs of local inflammation. The two smaller particle sizes (0.425 mm3 to 0.850 mm3 and 0.850 mm3 to 2.0 mm3) were minimally osteoinductive and were associated with a greater local inflammatory response. Microscopic events associated with new bone production in the two largest particle sizes included the presence of new osteoprogenitor cells in matrix vascular spaces, multinucleated osteoclast-like giant cells, and the formation of cartilage, osteoid, and new mineralized bone. The two smaller particle sizes were associated with increased fibrous tissue ingrowth and the presence of increased numbers of inflammatory cells. The temporal events of osteoinduction in response to heterotopic implantation of allogeneic DBM were delayed in horses when compared with other species. Although the demineralization technique used was effective in previous investigations on other species, incomplete demineralization occurred in the two smaller particle sizes that was not detected until after DBM implant harvest. The demineralization process may need to be altered for equine bone and precise monitoring for complete demineralization before implantation is required. Equine DBM is osteoinductive in muscle pouch sites in horses but matrix particle size does affect osteoinductive capability.