Clinical Evaluation of Demineralized Bone Matrix Allograft in Femur Fracture in Dogs

Eight clinical cases of dogs having femur fracture, were divided into two groups, A and B with four dogs (n=4) in each group. Group A animals were treated with intramedullary pinning (IMP) alone and group B were treated using IMP alongwith demineralized bone matrix (DBM) implantation at the fracture site. The efficacy of healing was evaluated on the basis of clinical evaluation, haemato-biochemical and radiographic parameters on the day of admission (day 0), followed by 7th, 15th, 30th, 45th and 60th post-operative days. The weight bearing was observed to be better in dogs of group A. Swelling was completely absent after 15th post-operative day in dogs of both the groups. The mean values of Hb, PCV, TLC and DLC showed a non-significant variation on subsequent post-operative days in both the groups. There was a significant increase in serum calcium up to 30th post-operative day and thereafter, followed a decreasing trend. The serum alkaline phosphatase values showed non-significant variation in group A while in group B animals significant increase observed on 15th, 30th and 45th post-operative day. Radiographically, the dogs of group B showed better radiographic union of fracture evidenced by early disappearance of fracture line than those of group A

Polyetheretherketone Cage with Demineralized Bone Matrix Can Replace Iliac Crest Autografts for Anterior Cervical Discectomy and Fusion in Subaxial Cervical Spine Injuries

OBJECTIVE: This study aimed to compare the clinical and radiologic outcomes of patients with subaxial cervical injury who underwent anterior cervical discectomy and fusion (ACDF) with autologous iliac bone graft or polyetheretherketone (PEEK) cages using demineralized bone matrix (DBM). METHODS: From January 2005 to December 2010, 70 patients who underwent one-level ACDF with plate fixation for post-traumatic subaxial cervical spinal injury in a single institution were retrospectively investigated. Autologous iliac crest grafts were used in 33 patients (Group I), whereas 37 patients underwent ACDF using a PEEK cage filled with DBM (Group II). Plain radiographs were used to assess bone fusion, interbody height (IBH), segmental angle (SA), overall cervical sagittal alignment (CSA, C2–7 angle), and development of adjacent segmental degeneration (ASD). Clinical outcome was assessed using a visual analog scale (VAS) for pain and Frankel grade. RESULTS: The mean follow-up duration for patients in Group I and Group II was 28.9 and 25.4 months, respectively. All patients from both groups achieved solid fusion during the follow-up period. The IBH and SA of the fused segment and CSA in Group II were better maintained during the follow-up period. Nine patients in Group I and two patients in Group II developed radiologic ASD. There were no statistically significant differences in the VAS score and Frankel grade between the groups. CONCLUSION: This study showed that PEEK cage filled with DBM, and plate fixation is at least as safe and effective as ACDF using autograft, with good maintenance of cervical alignment. With advantages such as no donor site morbidity and no graft-related complications, PEEK cage filled with DBM, and plate fixation provide a promising surgical option for treating traumatic subaxial cervical spine injuries.

Rehabilitation effect of BMSCs that was transfected with hBMP-2 through adenovirus combination with DBM on rabbit osteonecrosis in femoral head / 天津医药

Objective To evaluate the effect of human bone morphogenetic protein 2 (hBMP-2)/Bone Mesenchymal Stem Cells (BMSCs)/demineralized bone matrix(DBM) on repairing rabbits’femoral head after necrosis and to explore the new treatments for femoral head necrosis. Methods Femoral head necrosis models was established by clinical core decom?pression combined with liquid nitrogen frozen. Then, animals were randomly devided into 4 groups (n=12 per group):Group A were not implanted anything as control group, Group B were implanted with DBM. Group C were implanted with hBMP-2/DBM. Group D were implanted with hBMP-2/BMSCs/DBM. Four rabbits from each group were sacrificed at 4,8 and 12 weeks after surgery to evaluate the the repairing effect of Osteonecrosis of the femoral head (ONFH) through X-ray examina?tion, observation of the specimen and HE staining. Results X-ray revealed defect of femoral head in Group A without clear bone formation. There is a little fibrous hyperplasia and no obvious osteogenic response. By contrast, the femoral head defect areas became fuzzy in group B, group C and group D with new bone trabeculars. And the regenerate phenomenons of group D were significantly better than that of group B and group C of the same time point. As to the Lane-Sandhu X Ray scores, it is lower in group A than that in group B;It is lower in group C than that in group D(P<0.05). There is no statistical difference between Group B and Group C. General observation of the specimen revealed that the femoral head of group A collapsed with drilling holes. The femoral heads of group B and group C showed no collapse but the drilling holes existed. Femoral head in group D was not collapsed and the drilling holes disappeared. HE staining showed that bone trabeculars became ne?crotic and fragmented in Group A with a lot of air trapped cells. There were newborn immature bone trabeculars and osteo? blasts in group B and group C. Group D were of large number of bone cells, fat cells, and newborn mature bone trabeculars. The ratio of empty lacuna is higher in Group A than that in Group B;it is higher in Group C than that in Group D(P<0.05). Conclusion hBMP-2/BMSCs/DBM can induce BMSCs differentiation into osteoblasts after being implanted. It has good re?pairing effect on ONFH with good application prospect.

The behavior of demineralized bone matrix (DBM) in post-extraction sockets / El comportamiento de matriz ósea desmineralizada (DBM) en alvéolos post-extracción

Autogenous bone grafts are considered to be the gold standard in bone regeneration because of their osteogenic activity; however, due to limited availability of intraoral donor sites and the need to resolve the demands of patients requires an alternative to these. Two male patients were submitted to implant surgery in two stages with 6 months intervals between each of them: the first was exodontia and placement of DBM graft into the socket; the second stage was the drill with a 2 mm internal diameter trephine in center of the alveolar ridge previously grafted with DBM and subsequent implant placement. The samples were analyzed under histological techniques. A very mature bone was observed at 6 months after DBM graft placement in the sockets, showing it to be a good alternative as bone graft.

Los injertos de hueso autólogo son considerados el "gold standard" en regeneración ósea debido a sus propiedades osteogénicas; sin embargo, debido a la limitada cantidad de sitios intraorales y la necesidad de resolver las necesidades de los pacientes, es que se requiere una alternativa para el mismo. Dos pacientes masculinos fueron sometidos a una cirugía de implantes en dos etapas con 6 meses de intervalo entre cada una de ellas: la primera consistió en la exodoncia y el posicionamiento del injerto de DBM en el alvéolo; la segunda etapa consistió en el fresado con una trefina de dos milímetros de diámetro interno en el centro del reborde alveolar previamente injertado con DBM y luego se posicionó el implante. Las muestras fueron analizadas bajo técnicas histológicas. Un hueso muy maduro fue observado a los 6 meses de haber injertado el DBM en los alvéolos, demostrando que podría ser una buena alternativa como injerto óseo.

Mortality of Plutella xylostella (Lepidoptera, Plutellidae) by parasitoids in the Province of Santa Fe, Argentina

Mortality of Plutella xylostella (Lepidoptera, Plutellidae) by parasitoids in the Province of Santa Fe, Argentina. Plutella xylostella (Linnaeus, 1758) (Lepidoptera, Plutellidae) larvae cause severe economic damage on cabbage, Brassica oleracea L. variety capitata (Brassicaceae), in the horticultural fields in the Province of Santa Fe, Argentina. Overuse of broad spectrum insecticides affects the action of natural enemies of this insect on cabbage. The objectives of this work were to identify the parasitoids of P. xylostella and to determine their influence on larva and pupa mortality. Weekly collections of larvae and pupae were randomly conducted in cabbage crops during spring 2006 and 2007. The immature forms collected were classified according to their developmental stage: L1 and L2 (Ls = small larvae), L3 (Lm = medium larvae), L4 (Ll = large larvae), pre-pupae and pupae (P). Each individual was observed daily in the laboratory until the adult pest or parasitoid emergence. We identified parasitoids, the number of instar and the percentage of mortality of P. xylostella for each species of parasitoid. Parasitoids recorded were: Diadegma insulare (Cresson, 1875) (Hymenoptera, Ichneumonidae), Oomyzus sokolowskii (Kurdjumov, 1912) (Hymenoptera, Eulophidae), Cotesia plutellae (Kurdjumov, 1912) (Hymenoptera, Braconidae) and an unidentified species of Chalcididae (Hymenoptera). Besides parasitoids, an unidentified entomopathogenic fungus was also recorded in 2006 and 2007. In 2006, the most successful parasitoids were D. insulare and O. sokolowskii, while in 2007 only D. insulare exerted a satisfactory control and it attacked the early instars of the pest.

Mortalidade de Plutella xylostella (Lepidoptera, Plutellidae) por parasitóides na Província de Santa Fé, Argentina. Plutella xylostella (Linnaeus, 1758) (Lepidoptera, Plutellidae) causa danos econômicos severos em repolho, Brassica oleracea variedade capitata L. (Brassicaceae), na área de horticultura localizada na Província de Santa Fé, Argentina. O uso excessivo de inseticidas de largo espectro afeta a ação dos inimigos naturais de P. xylostella em repolho. O presente trabalho teve como objetivo identificar os parasitóides de P. xylostella e determinar sua influência na mortalidade de larvas e pupas em Santa Fé, Argentina. Coletas semanais de larvas e pupas em culturas de repolho foram realizadas aleatoriamente na área durante a primavera de 2006 e 2007, registradas pelo estágio de desenvolvimento: L1 e L2 (Ls = larvas pequenas), (Lm = larvas médias) L3, L4 (Ll = larvas grandes) pré-pupa e pupa (P). Cada indivíduo foi observado diariamente no laboratório até a emergência dos adultos da praga ou dos parasitóides. Foram identificadas as espécies de parasitóides o número de instars e a porcentagem de mortalidade. Os parasitóides registrados foram: Diadegma insulare (Cresson, 1875) (Hymenoptera, Ichneumonidae), Oomyzus sokolowskii (Kurdjumov, 1912) (Hymenoptera, Eulophidae), Cotesia plutellae (Kurdjumov, 1912) (Hymenoptera, Braconidae) e uma espécie não identificada pertencente à família Chalcididae. Além dos parasitóides, um fungo entomopatogênico não identificado foi registrado. Em 2006, os parasitóides mais bem sucedidos foram D. insulare e O. Sokolowskii; enquanto que em 2007 apenas D. insulare exerceu um controle satisfatório, atacando os instares iniciais da praga.

Effect of Curettage and DBM-CaSO4 Graft for the Treatment of Ischemic Necrosis of the Capital Femoral Epiphysis in Immature Pigs / 대한정형외과연구학회지

PURPOSE: The purpose of this study was to determine the effect of curettage and DBM complex graft as a new treatment modality for LCP disease using piglet capital femoral epiphysis ischemic necrosis model. MATERIALS AND METHODS: Five to six weeks old piglets were used for the experiment. Ischemic necrosis of the capital femoral epiphysis was surgically induced by cervical ligation on both sides. Three weeks following ischemic insult, the left hip joint was approached medially. About 15% of the necrotic capital femoral epiphysis was curetted through a window which was opened at medial cervical cortex, then, demineralized bone matrix complex was engrafted. The right femoral heads served as controls. Piglets were sacrificed three, six, nine, and twelve weeks following were harvested for histologic examination. RESULTS: In control group, photomicrographs of specimens showed central necrosis and fibrovascular invasion in capital femoral necrosis at three weeks after ischemic insult. Six, nine, and twelve weeks following ischemic insult, fibrovascular invasion advanced without noticeable new bone formation and collapse of femoral head progressed. At twelve weeks, definite coxa plana developed. In curettage and DBM complex graft group, there was evident new bone formation observed in the site of DBM complex graft. At three weeks, new bone formation along with fibrovascular invasion was observed around the engrafted DBM complex mainly in the cervical metaphyseal area. At six and nine weeks, new bone formation progressed into the engrafted DBM complex in the cervical metaphysis and around the engrafted DBM complex in the capital femoral epiphysis. At twelve weeks, new bone along with new cartilage formation was observed in the capital femoral epiphysis. CONCLUSION: In conclusion, curettage and DBM complex graft is thought to be an effective treatment modality that promote regeneration of ischemic necrosis of capital femoral epiphysis.

Efficiency of Anterior Interbody Fusion using Cage Packed with DBM in the Distractive Flexion Injury of Cervical Spine: Demineralized Bone Matrix vs Autoiliac Cancellous Bone

STUDY DESIGN: This is a retrospective study. OBJECTIVES: We wanted to evaluate the effectiveness and safety of a PEEK cage filled with DBM in patients with a distractive flexion injury of the cervical spine. SUMMARY OF LITERATURE REVIEW: AIF of the cervical spine using an autoiliac bone graft and plate fixation is known to be an effective treatment for traumatic injuries. However, the complications arising from the donor site are troublesome, and so fusion with cage is an alternative treatment. MATERIALS AND METHODS: We analyzed 32 cases (22 males and 10 females) with distractive flexion injury of the cervical spine. They underwent anterior decompression and interbody fusion with a PEEK cage and anterior plate fixation. In 18 patients, the cage was filled with autogenous iliac bone (Group I), and for the other 10 the cages were filled with DBM (Group II). RESULTS: All the cases in Group I and Group II achieved fusion except for one case of nonunion in group II. The anterior and posterior vertebral heights of the fused segments of group II were decreased more than those of group I, resulting a statistical difference (p=0.003). The changes of segmental lordosis (p=0.69) and the neurologic status (p=0.22) showed no statistical difference between the two groups. CONCLUSIONS: AIF using a PEEK cage filled with DBM and plate fixation showed no significant clinical differences compared to the case of iliac bone autografting. However, from a radiologic perspective, the time to achieve union was extensive and a case of nonunion was also observed. Therefore, many considerations are necessary when using DBM as a replacement for iliac bone autografting and further research should be done on this subject.

Structural augmentation with biomaterial loaded allograft threaded cage for the treatment of femoral head osteonecrosis surgical technique / 中国矫形外科杂志

[Objective]To introduce the surgical technique and its rationale and to evaluate the early and mid-term clinical outcome of routine core decompression and insertion of a biomaterial loaded allograft threaded cage(ATC)in the treatment of necrotic femoral head．[Methods]Seventy-six patients(78 hips)with femoral head necrosis were allocated to a program of either core decompression or core decompression and implantation of ATC．[Results]At review all patients had a minimum follow-up of 24-months(24 to 68 months)．In the control group，no significant improvement in Harris hip score(HHS)was found，and 13 of the 22 hips had deteriorated to stageⅢ．In the treatment group．the mean HHS was improved from 62．8 to 81．6．Collapse was seen in 1 hip，and this collapse Was progressive in 3 hips．[Conclusion]It，s thus evident that the technique is attractive as a salvage procedure．which shows encouraging Success rates and early clinical results.

Effect of Murine Adipose Derived Stem Cell(ADSC) on Bone Induction of Demineralized Bone Matrix(DBM) in a Rat Calvarian Defect Model

PURPOSE: Adipose tissue-derived stem cells(ADSC) has an osteoconductive potential and demineralized bone matrix(DBM) is an osteoinductive material. A combination of DBM and ADSC wound probably create osteoinductive properties. The purpose of this study is to determine the effect of the combination of DBM and ADSC mixture on healing of rat calvarial defect. METHODS: Thirty adult male Sprague-Dawley rats were randomized into 3 groups(n=10) as 1) Control, 2) DBM alone, 3) DBM with ADSC mixture. DBM with ADSC mixture group has had a 3-day preculture of ADSC from groin fat pad. An 6 mm critical size circular calvarial defect was made in each rat. Defect was implanted with DBM alone or DBM with ADSC mixture. Control defect was left unfilled. 6 and 12 weeks after the implantation, the rats were sacrificed and the defects were evaluated by histomorphometric and radiographical studies. RESULTS: Histomorphometric analysis revealed that DBM with ADSC mixture group showed significantly higher bone formation than DBM alone group(p<0.05). Although radiographs from DBM alone group and DBM with ADSC group revealed similar diffuse radiopaque spots dispersed throughout the defect. Densitometric analysis of calvarial defect revealed DBM with ADSC mixture group significantly higher bone formation than DBM alone(p<0.05). There was correlation of densitometry with new bone formation(Spearman's correlation of coefficient=0.804, 6 weeks, 0.802, 12 weeks). CONCLUSION: The DBM with ADSC mixture group showed the best healing response and the osteoinductive properties of DBM were accelerated with ADSC mixture. It will be clinically applicable that DBM and ADSC mixture in plastic and reconstructive surgery, such as alveolar cleft and congenital facial deformities that bone graft should be required.

Compatibility of Self-setting DBM-CP Composites in Percutaneous Kyphoplasty

PURPOSE: To analyze the physical properties of demineralized bone matrix (DBM) and self-setting calcium phosphate cement (CPC) composite for its compatibility to percutaneous kyphoplasty. MATERIALS AND METHODS: According to tap volume method, DBM was mixed with CPC in variable ratio 0%, 20%, 30%, 40% and 50%. Distilled water was used as a hardening fluid. Its properties, including injectability, mold applicability, setting time and its behavior, maximum temperature, and mechanical strength, were analyzed. RESULTS: The DBM-CP composites has a good injectability and mold applicability, a maximum temperature of less than 5oC, a initial setting time of 3 to 10 minutes. The outer surface of DBM-CP composites showed their even distribution in optical microscopy. Injectability, mold applicability and compressive strength were in inverse proportion to the amounts of DBM. CONCLUSION: This study suggests that the DBM-CP composites has a good injectability and mold applicability with a low setting temperature and even distribution of compound. Therefore this composite might be used as a substitute of PMMA in kyphoplasty.

Preparation of nano DBM and its properties / 中国矫形外科杂志

[Objective] To observe the structure characteristics of nano decalcified bone matrix(DBM)produced by MICROS super fine mill,and to study the nano-technologies of DBM and the biocompatibility of nano-DBM as bone graft substitutes.[Methods]Improved Urist method was used to produce DBM.The pieces of DBM was pre-porphyrizated by liquid nitrogen frozen ball mill equipment.The DBM powder were further porphyrizated by MICROS super fine mill to be nano-DBM.The structure of nano-DBM was observed by electron microscope.The experiments on acute toxicity,pyrogen and hemolysis were performed according to theTechnical Evaluation Standards of Biomedical Materials and Medical Instruments promulgated by Chinese Ministry of Health.[Results]The DBM was porphyrizated to the size of 50-200 nm in diameter.No toxicity,pyrogen nor hemolysis of nano-DBM was noted.Nano-DBM exhibited excellent biocompatibility.[Conclusion]Nano-DBM produced under the conditions of low or controlled temperature,is a kind of biomaterial which is avirulent,nonirritant,nonpyrogenic,nonimmunological reaction and reveals good biocompatibility.

The effects of cryopreservation on growth and osteogenesis of human bone marrow stromai cells cultured on demineralized bone matrix / 中华创伤骨科杂志

Objective To investigate the effects of cryopreservation on the growth and osteogenesis capa- bility of human bone marrow stromal cells(BMSCs)on demineralized bene matrix(DBM).Methods Bone marrow aspirates were obtained from the lilac crests of three donors.The BMSCs were isolated from the bone marrow by density gradient centrifugation.Cells of passage 3 were cryopreserved in liquid nitrogen for 24 hours,and then re- covered.The non-cryopreserved BMSCs were used as the control,The cryopreserved and control BMSCs were cul- tured in osteogenic media,collected and labeled with Dil to be seeded onto the DBM when cells were confluent.The percentage of BMSCs adhered to the DMB was detected.The cell morphology and matrices secreted by BMSCs on the DBM were observed by the inverted phase-contrasted microscope,fluorescence microscope and scanning electron microscope(SEM).The growth and viability of BMSCs on the DBM were determined using the modified MTT ashy. The osteogenesis ability of BMSCs on the DBM was determined by assessment of the alkaline phosphatase(ALP) activity and osteocalcin(OCN)content.Results The percentages of the cryopreserved and control cells adhered to DBM were(97.25?1.17)% and(97.00?1.09)% respectively.The cells adhered well to the DBM and grew rapidly.Large amounts of matrices on the DBM were observed by the light microscope and SEM.The cells embedded in the matrices could be observed by fluorescence microscope.There were no significant differences in the assay values of MTT,ALP and OCN between the cryopreserved and control BMSCs on the DBM.Conclusion Since cryopreservation does not affect the growth and osteogenesis capability of BMSCs on DBM,the cryopreserved BMSCs can be used as a cell source in bone tissue engineering.

Biocompatibility of PDLLA/HA/DBM with human osteoblasts co-cultured in vitro / 第三军医大学学报

Objective To evaluate the osteocompatibility of poly(D,L lactic acid)/ hydroxyapatite/decalcified bone matrix (PDLLA/HA/DBM) and to compare the osteocompatibility with that of PDLLA and DBM. Methods The isolated human osteoblasts from the femoral head of patients were inoculated onto PDLLA/HA/DBM, PLA, and DBM. The interface between biomaterial and osteoblasts was investigated with phase contrast microscope and scanning electron microscope. Results Osteoblasts were attached tightly to the surface of biomaterial with an arched structure and had normal morphology. The extracellular collagenous matrix covered the surface of biomaterial and packed the granules of biomaterial completely. Conclusion PDLLA/HA/DBM can form osteointegrated interface at the early stage with good biocompability.