Biomedical subjects

J O Hollinger

Publications and source records attributed to J O Hollinger.

At least 37 records · Page 2Linked to original sources

PubMed1998

Recombinant human bone morphogenetic protein-2 and collagen for bone regeneration.

The study reported describes a combination of recombinant human bone morphogenetic protein-2 (rhBMP-2) and collagen (C) to regenerate bone. Unilateral critical-sized defects (CSDs) were prepared in radii of 32 skeletally mature New Zealand white rabbits. Rabbits were divided evenly among four treatments: autograft, absorbable C (Helistat), 35 microg of rhBMP-2 combined with absorbable C (rhBMP-2/C), and untreated CSDs. The two euthanasia periods were 4 and 8 weeks. Radiographs were taken the day of surgery, every 2 weeks, and at term and the percent of radiopacity was measured. Data analysis revealed a time-dependent increase in the percent radiopacity with rhBMP-2/C. Histological examination revealed the rhBMP-2/C treatment regenerated osseous contour by 8 weeks. According to quantitative histomorphometry, the CSD and C groups had significantly less new bone than either autograft or rhBMP-2/C (p < or = 0.05). The results suggest that rhBMP-2/C could be an effective therapy to restore segmental bone defects.

Animals↗

PubMed1998

Radiomorphometry and biomechanical assessment of recombinant human bone morphogenetic protein 2 and polymer in rabbit radius ostectomy model.

The study objective was to determine the mechanical integrity and radiopacity of regenerated bone within critical-sized defects (CSDs) in radii of rabbits using recombinant human bone morphogenetic protein 2 (rhBMP-2) with a porous, biodegradable poly(D,L-lactic acid) (PDLLA) carrier (designated PLA). Twenty millimeter, unilateral radial ostectomies were created in 96 skeletally mature New Zealand white rabbits. The rabbits were randomly assigned to six treatment groups with two euthanasia periods. Treatment groups included unfilled defect (n = 8), segmental autograft (n = 8), PLA + 0 microg rhBMP-2 (n = 8), PLA + 17 microg rhBMP-2 (n = 8), PLA + 35 microg rhBMP-2 (n = 8), and PLA + 70 microg rhBMP-2 (n = 8). The radiopacity was significantly greater for the 35- and 70-microg rhBMP-2 groups at 4 weeks compared to unfilled controls, PLA only, and 17-microg rhBMP-2 groups and equivalent to the autograft. At 8 weeks all groups receiving rhBMP-2 were equivalent to the autograft and significantly greater than unfilled defects and PLA alone. Similarly, the biomechanical analysis indicated significantly greater torque at failure for the 35-microg rhBMP-2 group compared to all other groups at 4 weeks. By 8 weeks all groups receiving rhBMP-2 and autograft had significantly greater torque than unfilled controls and PLA alone. These radiomorphometric and biomechanical results indicate PLA may be a suitable carrier for rhBMP-2 used for skeletal regeneration.

Animals↗

PubMed1998

Isolation of human osteoblast-like cells and in vitro amplification for tissue engineering.

As the field of dental implants continues to grow at a rapid rate so does our quest to find new techniques to enhance bone grafting. Tissue engineering is an exciting new technique in bone grafting. Therefore, the purposes of this study were to develop a simple, reproducible method to isolate human osteoblast-like cells (HOBs) and to evaluate in vitro cell proliferation within 2 different 3-dimensional (3-D) constructs targeted for tissue engineering applications. Ultimately, HOBs that have been amplified within 3-D constructs may be employed for bone regeneration techniques, such as onlay and sinus grafting prior to implant placement. Our cell isolation protocol employed human fetal calvaria tissue sequentially digested with trypsin and collagenase. The HOB cells from only the third and fourth digests were obtained, cultured and evaluated within the constructs. An osteoblast-like phenotype was in part verified for these HOB cells by demonstrating a significantly higher alkaline phosphatase activity than for human gingival fibroblasts, and a comparable level to the osteoblast cell line MG-63. The HOB cells were cultured within either poly (D,L-lactide) (PLA) or a fused fiber ceramic and evaluated for the ability to support in vitro HOB amplification. HOB proliferation was validated by scanning electron microscopy, identifying cells throughout the 3-D constructs. Continuous cell viability was demonstrated for the duration of the 33-day evaluation period and the extent of cell amplification reached approximately 20 times the seeding density. The in vitro amplification results further indicate that tissue engineering strategies with either the PLA or fused fiber construct may be suitable for bone regeneration therapy for dental implants.

Biomedical Engineering↗

PubMed1997

Macrophysiologic roles of a delivery system for vulnerary factors needed for bone regeneration.

Traditional histology identifies three components of bone: cells, an extracellular mineralized organic matrix, and a lymphatic-vascular component. Specialized bone cells known as osteoblasts promote bone regeneration. Clinically, this property has been exploited by surgeons with autografts and bank bone preparations to restore deficient form and function to almost every aspect of the skeleton. Unfortunately, these therapies can be inadequate for patients with panskeletal trauma. Therefore, a suitable alternative may be a laboratory-derived product consisting of a vulnerary factor and delivery system. The integration of a laboratory-engineered product in an osseous wound environment is a formidable challenge demanding a keen appreciation of the product's macrophysiologic roles in wound healing biology. Consequently, the purposes for this paper are 1) to define briefly macrophysiology relevant to a delivery system for vulnerary molecules and bone regeneration; 2) to review a key family of bone regenerating molecules, the bone morphogenetic proteins (BMPs); and 3) to relate delivery system engineering with bone regeneration.

Animals↗

PubMed1997

Evaluation of particulate Bioglass in a rabbit radius ostectomy model.

Osseous defects and fractures may require supplimentation to support and promote healing. Bioglass (BG) may be a useful therapeutic for these conditions. Therefore, we executed a study to determine whether particulate BG could promote healing of 20-mm unilateral ostectomies in the radius of rabbits. Ostectomies were either treated with BG or remained untreated in the control (CTL) group. At 4 and 8 weeks post-treatment, ostectomies were assessed histomorphometrically and biomechanically. New bone formation was more intense contiguous to the host bone for both BG and CTL than centrally, yet BG animals displayed active mineralization throughout the ostectomy. The amount of bone within BG-filled defects was greater than CTLs at 4 weeks, whereas, at 8 weeks there was no difference. Biomechanically, the BG-treated limbs required more torque to break than did CTL limbs at 4 weeks; however differences were not significantly different. By 8 weeks, the BG-treated and CTLs, had comparable strength. Bioglass may be a useful therapy to produce the early phase of osseous repair. However, improvements in handling properties of the particles will be needed to enhance efficacy.

Animals↗

PubMed1997

Comparison of porous bone mineral and biologically active glass in critical-sized defects.

Several materials have been proposed as therapies to augment alveolar bone and to promote periodontal regeneration. However, there are an insufficient number of studies that effectively evaluated these therapies. Consequently, the purpose of this study was to compare bone regeneration promoted by porous bone mineral and biologically active glass. Unilateral critical-sized defects (CSDs) were prepared in the radii of 24 rabbits, divided evenly between 2 time periods (4 and 8 weeks) and between 2 treatment groups (porous bone mineral and biologically active glass). Evaluations consisted of clinical examinations, standardized radiography at baseline and every 2 weeks thereafter, as well as histology and histomorphometry. Data were analyzed by an unpaired Student t-test with significance established at P < or = 0.05. We determined that CSDs treated with porous bone mineral were significantly more radiopaque than biologically active glass-treated sites at both 4 and 8 weeks. Moreover, the amount of new bone was significantly greater at both 4 and 8 weeks in the porous bone mineral groups than in the biologically active glass groups. We concluded that in the rabbit radius CSD wound model, porous bone mineral appears to be more effective than biologically active glass in regenerating bone.

Alveolar Bone Loss↗

PubMed1997

Bone formation with use of rhBMP-2 (recombinant human bone morphogenetic protein-2).

We examined the effect of rhBMP-2 (recombinant human bone morphogenetic protein-2), delivered in a porous poly(DL-lactic acid) implant, on bone formation in a critical-sized defect in the radial diaphysis in rabbits. A unilateral segmental defect, twenty millimeters long, was created in the radius in ninety-six skeletally mature New Zealand White rabbits. Forty-eight rabbits were evaluated at four weeks and forty-eight, at eight weeks. Six groups were studied at each time-period. The defect was left empty in one group (control), the defect was filled with an autogenous corticocancellous bone graft in one group, and the defect was filled with a porous poly(DL-lactic acid) implant containing zero, seventeen, thirty-five, or seventy micrograms of rhBMP-2 (one group each). Radiographs of the defects were made every two weeks. The percentage of the total area of the defect that was radiopaque was determined with use of computerized radiomorphometry, and this percentage was used as a quantitative measure of the extent of new-bone formation in the defect. There were time and dose-dependent responses to rhBMP-2 for as long as four weeks; thereafter, the effects of seventeen, thirty-five, and seventy micrograms of rhBMP-2 were independent of dose and time (p < or = 0.05). The defects that had been treated with either thirty-five or seventy micrograms of rhBMP-2 had a significantly greater (p < or = 0.05) area of radiopacity than the defects that had been treated with either zero or seventeen micrograms of rhBMP-2. No significant difference could be found between the defects treated with thirty-five or seventy micrograms of rhBMP-2 and the defects filled with an autogenous graft. Healing and bone formation were examined histologically and histomorphometrically as well. At four weeks, polarized light microscopy revealed remnants of poly(DL-lactic acid) only in the defects that had been filled with an implant containing zero micrograms of rhBMP-2. At eight weeks, regardless of the dose of rhBMP-2, poly(DL-lactic acid) was not visible on histological examination. The presence of multinucleated giant cells was the hallmark of the inflammatory response elicited by poly(DL-lactic acid). At four and eight weeks, macrophages and lymphocytes were also present. The intensity of the cellular response at four weeks suggested an inverse relationship between these cells and the dose of rhBMP-2 -- that is, there appeared to be more multinucleated giant cells in defects treated with zero micrograms of rhBMP-2 than in defects treated with seventy micrograms of rhBMP-2. At eight weeks, multinucleated giant cells were rare in the defects treated with seventeen, thirty-five, or seventy micrograms of rhBMP-2. Histomorphometric data at four and eight weeks indicated that the amount of bone formation in the defects treated with seventeen, thirty-five, or seventy micrograms of rhBMP-2 was equivalent to the amount in the defects treated with an autogenous graft and was significantly less (p < or = 0.05) in the untreated defects and the defects treated with zero micrograms of rhBMP-2 (p < or = 0.05). By eight weeks, only thirty-five and seventy micrograms of rhBMP-2 had restored cortices and marrow elements.

Animals↗

PubMed1996

Poly(alpha-hydroxy acids): carriers for bone morphogenetic proteins.

A broad spectrum of cells and cell products is associated with bone homeostasis and the renewal of bone following injury. The coupled interactions among cells provide the power behind sculpting of bone, sustaining form, and ensuring functionality. Local and systemic regulatory molecules (e.g. growth factors, hormones) direct cellular interactions through autocrine, paracrine, and hormonal pathways. Recently, genes for a class of osteogenic regulatory molecules have been cloned, and gene product expression has enabled investigators to assess safety and efficacy in animal studies. The molecules are known as bone morphogenetic proteins (BMPs). Therapeutic applications of BMPs depend on a carrier system. A carrier could spatially and temporally localize BMP for regional needs and be custom-tailored for acute craniofacial applications or for recalcitrant extremity non-unions. The poly(alpha-hydroxy acids) (PHAs) may be suitable for these applications. Therefore, the purposes of this paper are (i) to mention, briefly, basic concepts of the bone wound continuum and the possible therapeutic roles of BMPs; (ii) to outline several properties of selected PHAs relevant to bone regeneration dynamics; and (iii) to review selected preclinical studies with PHAs.

Animals↗

PubMed1996

Integrated processes responsible for soft tissue healing.

Wounded soft tissues undergo repair through a complex series of interrelated events that involve both physical and chemical activities. These processes are currently undergoing extensive investigation as efforts are directed toward achieving augmented and accelerated healing. Early wound-healing research focused on expanding traditional histologic descriptions of tissue healing by attempting to characterize the environment and biologic mediators responsible for healing. These initial studies successfully identified a number of agents and physiochemical factors present in healing wounds, but their precise roles and importance remain largely unknown. This review article summarizes the current literature on soft tissue healing. An effort has been made to correlate the activities of the major growth factors and cytokines with the individual reparative processes including the inflammatory response, hemostasis, fibroplasia, angiogenesis, and remodeling. Explanations and characteristics of growth factor function as well as brief descriptions of several major factors and their spectrum of activity are also provided.

Blood Coagulation↗

PubMed1996

Initial biocompatibility studies of a novel degradable polymeric bone substitute that hardens in situ.

Clinical management of osseous defects often requires bone grafts. The standard for treatment is autogenous bone harvested from sites such as either the iliac crest or the outer table of the calvaria. In addition to the problem of donor site morbidity and the limited supply of graft material, there is the additional operating time associated with harvesting procedures. A synthetic, bone graft substitute that can match the clinical performance of autogenous bone could alleviate these deficiencies. Therefore, a polymeric bone substitute was developed that consists of a four-armed star polymer of poly(dioxanone-co-glycolide) endcapped at each termini with a biocompatible lysine-based diisocyanate crosslinker. The polymer can be mixed with inorganic fillers such as either hydroxyapatite or tricalcium phosphate to form either injectable or moldable putty. The addition of a catalyst (for example, diethylaminoethanol and water) to the polymer produces a crosslinking reaction causing the combination to harden. This reaction is nontoxic, normo-thermic and can be performed in situ. During the course of the polymerization, carbon dioxide is liberated, producing an interconnected porous network within the implant, suitable for bone ingrowth. This paper will describe a preliminary biocompatibility assay of the bone substitute.

Animals↗

PubMed1996

Role of bone substitutes.

Approximately 500 million years ago, the Paleozoic era heralded an evolutionary marvel: the skeleton. Unique to this evolutionary development was the capacity for regeneration: the physiologic renewal of embryologically derived tissue. Many of the cellular and molecular components for bone regeneration have been identified (bone morphogenetic proteins), and their therapeutic manipulation will become common clinical practice. Moreover, synthetic materials produced in the laboratory and novel bone derivatives will be used to exploit the skeleton's capacity to regenerate and repair. The concept of repair may be viewed as the restoration of form and function to deficient osseous tissue. Materials that provoke repair can be categorized broadly as bone substitutes. In this review, bone substitutes are grouped into 2 categories, polymers and ceramics, and each is subclassified as biodegradable or nonbiodegradable. Examples of these materials are provided as well as some of their liabilities and virtues.

Animals↗

PubMed1996

Onlay bone augmentation with an osteoinductive implant.

The repair of contour defects of craniofacial bone can be accomplished by several methods, including autogenous bone grafts, demineralized bone, and alloplastic materials. The objective of this study was to assess a biodegradable, xenogeneic, osteoinductive implant for craniofacial onlay bone augmentation. Twelve New Zealand White rabbits each had craniofacial onlays consisting of three experimental materials: 1) autograft; 2) allogeneic, demineralized block implant; and 3) partially purified osteoinductive protein (osteogenin) with allogeneic collagen and 50:50 poly (DL-lactide-co-glycolide). Implants with host bone were recovered after 20 weeks and assessed by histomorphometric methods. There was no evidence of adverse local reaction to the three treatments. No significant difference in bone replacement or bone density resulting from each of the implant types was demonstrated. In conclusion, the osteoinductive implants were well tolerated, and over the 20-week period they appeared to maintain their contour as onlays.

Animals↗

PubMed1996

A bone regeneration study: transforming growth factor-beta 1 and its delivery.

Critical-sized defects (CSDs) were prepared in 60 rabbit skulls, divided evenly among five treatments and two time periods. The treatments consisted of rabbit demineralized bone matrix (DBM) and four different doses of transforming growth factor-beta 1 (TGF-beta 1) (0, 0.4, 4, and 40 micrograms) delivered in 3% carboxymethyl cellulose and Gelfilm. Quantitative assessments of bone formation in the CSDs were accomplished with computerized radiomorphometry and histomorphometry. Results indicated that rabbit DBM and 40 micrograms TGF-beta 1 promoted more new bone formation at 4 weeks than the other treatments. By 8 weeks, bone formation in the CSDs was equivalent for all but the 0-microgram dose of TGF-beta 1. The results from the study suggest a single therapeutic dose of TGF-beta 1 delivered in an appropriate carrier to a critical-sized intraosseous wound should promote bone formation.

Analysis of Variance↗

PubMed1995

Incorporation of polylactide-polyglycolide in a cortical defect: neoangiogenesis and blood supply in a bone chamber.

Erodible polymers are an alternative to metals for fracture fixation (for example, in the malleolus) and for maxillofacial reconstruction. In this study, the vascular response to eroding polylactide-polyglycolide copolymer threads was observed chronically in a bone chamber implant, with use of intravital microscopy. A bone chamber implant loaded with 100 microns thick polylactide-polyglycolide threads was implanted into the right tibia in 15 mature female New Zealand White rabbits. Periodic intravital microscopic observations were performed from the third to the tenth or twelfth week after implantation. Vascularization, blood flow, and trabecular growth into the chambers from the medial cortex were recorded on videotape and analyzed using digital image processing. A statistically significant delay of neo-osteogenesis in the presence of this copolymer was described in an earlier report. The present report describes the measures of neoangiogenesis and blood supply; there was a significant delay in neoangiogenesis. It is suggested that both delayed angiogenesis and osteogenesis were secondary consequences of the macrophage response to slowly eroding poly-L-lactide crystal nanoparticles and the influence of reduced nutrient exchange. The lesser effect on blood supply and vascular volume fraction was seen to be linked to the slowing down of angiogenesis, as the latter allowed vessels to mature, with a widening of their calibers. This homeostatic adjustment was interpreted as being only partially successful in restoring control levels of oxygen delivery, because resulting increases in vessel surface area did not reach control levels. Thus, in the presence of eroding polylactide-polyglycolide, the oxygen supply and extravasation of other nutrients may be below normal during healing phases when the need is critical.

Analysis of Variance↗

PubMed1995

Calvarial bone repair with porous D,L-polylactide.

Bone is a storehouse of biologic factors enabling it to regenerate without scar formation. Recombinant technology has made many of these factors available in significant quantity for therapeutic applications. However, a system to deliver recombinant bone-regenerating factors is needed. Biodegradable, biocompatible polymers have shown promise for delivering bone regenerative factors, such as bone morphogenetic protein. The polymer we selected to investigate was racemic D,L-polylactide. Our immediate objective was to engineer porous D,L-polylactide to promote bone ingrowth (osteoconduction). We tested the hypothesis that porous D,L-polylactide implanted in a standard intraosseous calvarial wound would not hinder but would support bone regeneration. Therefore porous polylactide disks (65% void volume) were manufactured with pores < or = 100 microns, < or = 200 microns, and < or = 350 microns; implanted in rabbits' calvariae, and retrieved 1, 2, 4, and 6 months after insertion. Quantitative histomorphometry revealed a possible relationship in the amount of bone ingrowth with increasing pore size over time. The D,L-polylactide disks < or = 350 microns had the greatest quantity of bone ingrowth (< or = 0.05). However, a disturbing finding was the multinucleated giant cell response associated with all implanted disks. We speculate these cells may have produced an inhospitable environment stifling osteoconduction. Consequently, postsynthesis engineering refinements of D,L-polylactide to eliminate the giant cell response are crucial before loading with bone morphogenetic protein.

Animals↗

PubMed1994

Recombinant human bone morphogenetic protein-2 is superior to demineralized bone matrix in repairing craniotomy defects in rats.

The purpose of this study was to measure bone-regenerative effects of recombinant human bone morphogenetic protein-2 (rhBMP-2) in rat calvarial critical-size defects (CSDs). CSDs (8 mm in diameter) were treated with either 1) 2.2 micrograms rhBMP-2 combined with insoluble collagenous bone matrix (ICBM), 2) 6.5 micrograms rhBMP-2 plus ICBM, 3) ICBM alone, or 4) demineralized bone matrix (DBM), for 7, 14, or 21 days. Multiple linear regression showed that rhBMP-2 had a significant time- and dose-dependent effect on bone regeneration (P < .05). After 7 days, new calcifying cartilage and remineralizing ICBM, with an occasional zone of new woven bone, was evident in defects treated with rhBMP-2/ICBM. By 14 days, both doses of rhBMP-2 reconstituted with ICBM had induced more bone formation than ICBM alone or DBM, and 6.5 micrograms was superior to 2.2 micrograms. There was no evidence of adverse cellular response. This study shows for the first time that rhBMP-2 could restore osseous form to a calvarial defect. In addition, osteoregeneration was accelerated by the higher dose of rhBMP-2.

Animals↗

PubMed1994

Osseous regeneration in the rat calvarium using novel delivery systems for recombinant human bone morphogenetic protein-2 (rhBMP-2).

In the current investigation, we report osseous regeneration in critical-size rat calvarial defects using recombinant human bone morphogenetic protein-2 (rhBMP-2) and novel delivery systems based on biomaterials. The novel systems combine rhBMP-2 with dry powder microparticles of poly(D,L-lactide-co-glycolide) (PLGA). The mixture of rhBMP-2 with PLGA microparticles is added to an aqueous solution of biopolymer to yield a semisolid paste. The biopolymers tested include autologous blood clot, hydroxypropyl methylcellulose, and sodium alginate cross-linked with calcium ion. Insoluble collageneous bone matrix was also studied as a control. Test articles were made at 0-, 10-, and 30-micrograms doses of rhBMP-2 and imiplanted in 8-mm-diameter rat calvarial defects (which will not heal if left untreated). The animals were examined 21 days after implantation by radiography, radiomorphometry, histology, and histomorphometry. All tested materials containing rhBMP-2 restored radiopacity and normal contouring to the calvarial defects. Samples without added rhBMP-2 yielded only soft tissue within the defects. Histology showed restoration of inner and outer bone tables plus marrow constituents. The PLGA microparticles were significantly resorbed at the 21-day time point. Although small differences between delivery systems were evident at 0- and 10-micrograms rhBMP-2 doses, all test articles performed essentially equivalently at the 30-micrograms dose. Thus, novel delivery systems for rhBMP-2 offer the promise of combining the intrinsic bioactivity of the osteoinductive protein with pharmaceutically acceptable biomaterials.

Alginates↗

PubMed1994

Quantitative light microscopy. A powerful tool to assess bone.

Quantitative light microscopy is an important tool for assessing objectively the status of histologic events. It is, therefore, extremely useful for studying the dynamics of bone. This article provides a brief description of three quantitative techniques that are applicable to histomorphometry.

Animals↗