A comparison of the effects of ibuprofen and rofecoxib on rabbit fibula osteotomy healing.

BACKGROUND AND PURPOSE: Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit cyclooxygenase (COX) activity, which is the rate-limiting enzyme in the synthesis of prostaglandins. Previous studies have indicated that NSAID therapy, and in particular NSAIDs that specifically target the inflammatory cyclooxygenase (COX-2), impair bone healing. We compared the effects of ibuprofen and rofecoxib on fibula osteotomy healing in rabbits to determine whether nominal, continuous inhibition of COX-2 with rofecoxib would differentially affect fracture healing more than cyclical inhibition of COX-2 using ibuprofen, which inhibits COX-1 and COX-2 and has a short half-life in vivo. METHODS: Bilateral fibula osteotomies were done in 67 skeletally mature male New Zealand white rabbits. The rabbits were treated with placebo, rofecoxib (12.5 mg once a day), or ibuprofen (50 mg 3 times a day) for 28 days after surgery. Plasma ibuprofen levels were measured by HPLC analysis. Bone healing was assessed by histomorphometry at 3 and 6 weeks after osteotomy, and at 6 and 12 weeks by torsional mechanical testing. RESULTS: Plasma ibuprofen levels peaked and declined between successive doses. Fracture callus morphology was abnormal in the rofecoxib-treated rabbits and torsional mechanical testing showed that fracture healing was impaired. Ibuprofen treatment caused persistence of cartilage within the fracture callus and reduced peak torque at 6 weeks after osteotomy as compared to the fibulas from the placebo-treated rabbits. In the specimens allowed to progress to possible healing, non-union was seen in 5 of the 26 fibulas from the rofecoxib-treated animals as compared to 1 of 24 in the placebo group and 1 of 30 in the ibuprofen treatment group. INTERPRETATION: Continuous COX-2 inhibition as modeled by rofecoxib treatment appears to be more deleterious to fracture repair than cyclical cyclooxygenase inhibition as modeled by ibuprofen treatment. Ibuprofen treatment appeared to delay bone healing based upon the persistence of cartilage within the fracture callus and diminished shear modulus. Despite the ibuprofen-induced delay, rofecoxib treatment produced worse fracture (osteotomy) healing than ibuprofen treatment.

In vivo bone response to 3D periodic hydroxyapatite scaffolds assembled by direct ink writing.

The in vivo bone response of 3D periodic hydroxyapatite (HA) scaffolds is investigated. Two groups of HA scaffolds (11 mm diameter x 3.5 mm thick) are fabricated by direct-write assembly of a concentrated HA ink. The scaffolds consist of cylindrical rods periodically arranged into four quadrants with varying separation distances between rods. In the first group, HA rods (250 microm in diameter) are patterned to create pore channels, whose areal dimensions are 250 x 250 microm(2) in quadrant 1, 250 x 500 microm(2) in quadrants 2 and 4, and 500 x 500 microm(2) in quadrant 3. In the second group, HA rods (400 microm in diameter) are patterned to create pore channels, whose areal dimensions of 500 x 500 microm(2) in quadrant 1, 500 x 750 microm(2) in quadrants 2 and 4, and 750 x 750 microm(2) in quadrant 3. Each group of scaffolds is partially densified by sintering at 1200 degrees C prior to being implanted bilaterally in trephine defects of skeletally mature New Zealand White rabbits. Their tissue response is evaluated at 8 and 16 weeks using micro-computed tomography, histology, and scanning electron microscopy. New trabecular bone is conducted rapidly and efficiently across substantial distances within these patterned 3D HA scaffolds. Our observations suggest that HA rods are first coated with a layer of new bone followed by subsequent scaffold infilling via outward and inward radial growth of the coated regions. Direct-write assembly of 3D periodic scaffolds composed of micro-porous HA rods arrayed to produce macro-pores that are size-matched to trabecular bone may represent an optimal strategy for bone repair and replacement structures.

The effects of local platelet rich plasma delivery on diabetic fracture healing.

Several studies have documented that diabetes impairs bone healing clinically and experimentally. The percutaneous delivery of platelet rich plasma (PRP) was used in the diabetic BB Wistar femur fracture model to investigate the use of PRP as a concentrated source of critical early growth factors on bone healing. PRP delivery at the fracture site normalized the early (cellular proliferation and chondrogenesis) parameters while improving the late (mechanical strength) parameters of diabetic fracture healing. These results suggest a role for PRP in mediating diabetic fracture healing and potentially other high risk fractures.

The effects of local insulin delivery on diabetic fracture healing.

Several studies have documented that diabetes impairs bone healing clinically and experimentally. Systemic insulin treatment has been shown to ameliorate impaired diabetic bone healing. However, these studies failed to distinguish between a direct and a systemic effect of insulin upon bone healing. A novel intramedullary insulin delivery system was used in the diabetic BB Wistar femur fracture model to investigate the potential direct effects of insulin on bone healing. Insulin delivery at the fracture site normalized the early (cellular proliferation and chondrogenesis) and late (mineralized tissue, cartilage content and mechanical strength) parameters of diabetic fracture healing without affecting the systemic parameters of blood glucose. These results suggest a critical role for insulin in directly mediating fracture healing and that decreased systemic insulin levels in the diabetic state lead to reduced localized insulin levels at fracture site with concomitant increases in diabetic fracture healing time.

Mechanical testing of a novel hydrogel nucleus replacement implant.

BACKGROUND CONTEXT: Both total disc and nucleus replacement arthroplastic approaches are emerging as viable treatment modalities for discogenic pain resulting from degenerative disc disease. The results of the testing in this investigation show that this novel nucleus replacement implant exhibits physical and mechanical properties which make it uniquely suitable as a replacement to the degenerated nucleus pulposus. PURPOSE: The purpose of this investigation was to evaluate key performance characteristics of a novel nucleus replacement device. STUDY DESIGN: Mechanical tests were performed to evaluate essential properties of the implant. Of key importance was the ability of the implant to withstand fatigue testing and resist extrusion in cadaveric motion segments. Resistance of the implant to radial deformation is also relevant to the ability of the implant to resist extrusion and maintain disc height. METHODS: Resistance to radial bulging was demonstrated by applying axial compressive loads while examining the "footprint" of the implant through a clear plexiglass lower platen. A NeuDisc implant was compared with an isotropic implant produced from the same hydrolyzed polyacrylonitrile hydrogel. Confined compression tests were used to determine "lifting force" when implants were loaded within stainless steel cylinders submerged in Hanks' balanced salt solution. Fatigue tests involved loading for 10 million cycles. Implants were then examined for physical damage, retention of mechanical properties, and for the ability of the device to return to a complete level of hydration. The final evaluation was extrusion testing in a cadaveric model. Implanted specimens were either tested in compression, lateral bending, or flexion until catastrophic failure was apparent. RESULTS: The NeuDisc implant showed considerably less "bulging" when compared with the identically sized isotropic device. Confined compression testing revealed a lifting force of 400 N at 70% hydration for these hydrogel implants. After fatigue testing, all implants were physically intact and possessed similar swelling characteristics to control samples. In cadaver testing, mean force to failure in compression was 3581.3+/-1558.5 N. Mean moment to failure in lateral bending was 25.6+/-11.1 Nm. In flexion tests, the mean moment to failure was 52.2+/-18.3 Nm. End plate fracture and ligament failure were the most commonly observed failure modes. CONCLUSIONS: These initial results suggest that this novel implant may prove to be a suitable nucleus pulposus substitute.

The effects of blood glucose control upon fracture healing in the BB Wistar rat with diabetes mellitus.

Several clinical series, analyzing fracture healing in patients with diabetes mellitus (DM). demonstrated significant incidence of delayed union, non-union, and pseudarthrosis. In this study, analysis was performed to evaluate the effects of blood glucose (BG) control on fracture healing in the DM BB Wistar rat, a rat strain that represents a close homology to Type I DM in man. Our study showed decreased cell proliferation at the fracture site as well as decreased mechanical stiffness and bony content in the poorly controlled DM rats. To determine the effect of BG control, DM rats were treated with insulin sufficient to maintain physiologic BG levels throughout the course of the study. Values of cellular proliferation, biomechanical properties and callus bone content in tightly controlled DM animals were not significantly different from values of non-DM control values. This study suggests that insulin treatment with resultant improved BG control will ameliorate the impaired early and late parameters of DM fracture healing.

Low-intensity pulsed ultrasound increases the fracture callus strength in diabetic BB Wistar rats but does not affect cellular proliferation.

Type I diabetes mellitus (DM) is associated with impaired fracture healing. Specifically, DM affects multiple phases of fracture healing including early cellular proliferation and late phases resulting in inferior biomechanical properties. Recent studies demonstrated the utility of pulsed low-intensity ultrasound (US) to facilitate fracture healing. The current study evaluated the effects of daily application of US on mid-diaphyseal femoral fractures in DM and non-DM BB Wistar rats. Immunohistochemical staining for PCNA was used to evaluate cellular proliferation at 2, 4, and 7 days post-fracture. In concordance with previous findings. DM fracture callus demonstrated decreased cellular proliferation. Importantly, the application of US did not significantly alter the proliferation in either DM or control groups. However, mechanical testing revealed significantly greater torque to failure and stiffness in US-treated DM versus non-US-treated DM groups at six weeks post-fracture. Despite the inability of US to affect the early proliferative phase of fracture healing, its application clearly results in improved mechanical properties during the late phases of healing. These findings suggest a potential role of US as an adjunct for DM fracture treatment.

In vivo evaluation of a biomimetic apatite coating grown on titanium surfaces.

Osteoconductive mineral coatings represent an established technology for enhancing the integration of orthopedic implants with living bone. However, current coatings have limitations related to fabrication methods, attachment strength to metal substrates, and in vivo performance. Low temperature biomimetic growth is a coating technique wherein the device to be coated is immersed in a meta-stable saturated solution of the coating constituents and growth of the coating is then allowed to proceed on the surface of the device. This study focused on the in vivo evaluation of a biomimetic apatite coating fabricated under these conditions. The experiment was designed to specifically test the amount of bone ingrowth into the coated channels versus the uncoated channels of an established bone chamber system, with emphasis placed on the amount of bone present on the coupon surface. Three types of measurements were taken on each channel: linear ingrowth %, area ingrowth %, and continuous bone apposition %. The experiments demonstrated that under controlled conditions, the apatite coating appears to resorb in 8 weeks and did stimulate early osseointegration with the metal surface with a reduction in fibrous tissue encapsulation. This coating may, therefore, be useful in facilitating early bone ingrowth into porous surfaces without the potential for coating debris, macrophage infiltration, fibrous tissue encapsulation, and eventual coating failure as may occur with current plasma-sprayed hydroxapatite coating techniques.

Performance of hydroxyapatite bone repair scaffolds created via three-dimensional fabrication techniques.

The current study analyzes the in vivo performance of porous sintered hydroxyapatite (HA) bone repair scaffolds fabricated using the TheriForm solid freeform fabrication process. Porous HA scaffolds with engineered macroscopic channels had a significantly higher percentage of new bone area compared with porous HA scaffolds without channels in a rabbit calvarial defect model at an 8-week time point. An unexpected finding was the unusually large amount of new bone within the base material structure, which contained pores less than 20 microm in size. Compared with composite scaffolds of 80% polylactic-co-glycolic acid and 20% beta-tricalcium phosphate with the same macroscopic architecture as evaluated in a previous study, the porous HA scaffolds with channels had a significantly higher percentage of new bone area. Therefore, the current study indicates that scaffold geometry, as determined by the fabrication process, can enhance the ability of a ceramic material to accelerate healing of calvarial defects.

Engineered cellular response to scaffold architecture in a rabbit trephine defect.

Tight control of pore architecture in porous scaffolds for bone repair is critical for a fully elucidated tissue response. Solid freeform fabrication (SFF) enables construction of scaffolds with tightly controlled pore architecture. Four types of porous scaffolds were constructed using SFF and evaluated in an 8-mm rabbit trephine defect at 8 and 16 weeks (n = 6): a lactide/glycolide (50:50) copolymer scaffold with 20% w/w tri-calcium phosphate and random porous architecture (Group 1); another identical design made from poly(desaminotyrosyl-tyrosine ethyl ester carbonate) [poly(DTE carbonate)], a tyrosine-derived pseudo-polyamino acid (Group 2); and two poly(DTE carbonate) scaffolds containing 500 microm pores separated by 500-microm thick walls, one type with solid walls (Group 3), and one type with microporous walls (Group 4). A commercially available coralline scaffold (Interpore) with a 486-microm average pore size and empty defects were used as controls. There was no significant difference in the overall amount of bone ingrowth in any of the devices, as found by radiographic analysis, but patterns of bone formation matched the morphology of the scaffold. These results suggest that controlled scaffold architecture can be superimposed on biomaterial composition to design and construct scaffolds with improved fill time.

Performance of degradable composite bone repair products made via three-dimensional fabrication techniques.

This study analyzed the in vivo performance of composite degradable bone repair products fabricated using the TheriForm process, a solid freeform fabrication (SFF) technique, in a rabbit calvarial defect model at 8 weeks. Scaffolds were composed of polylactic-co-glycolic acid (PLGA) polymer with 20% w/w beta-tricalcium phosphate (beta-TCP) ceramic with engineered macroscopic channels, a controlled porosity gradient, and a controlled pore size for promotion of new bone ingrowth. Scaffolds with engineered macroscopic channels and a porosity gradient had higher percentages of new bone area compared to scaffolds without engineered channels. These scaffolds also had higher percentages of new bone area compared to unfilled control defects, suggesting that scaffold material and design combinations could be tailored to facilitate filling of bony defects. This proof-of-concept study demonstrated that channel size, porosity, and pore size can be controlled and used to influence new bone formation and calvarial defect healing.

Recombinant human platelet-derived growth factor BB (rhPDGF-BB) and beta-tricalcium phosphate/collagen matrix enhance fracture healing in a diabetic rat model.

Diabetes mellitus is a common systemic disease that has been associated with poor fracture healing outcomes. The mechanism through which diabetes impairs bone regeneration is unknown. One possible mechanism may be related to either decreased or uncoordinated release of local growth factors at the fracture site. Indeed, previous studies have found reduced platelet-derived growth factor (PDGF) levels in the fracture callus of diabetic rats, suggesting that local application of PDGF may overcome the negative effects of diabetes and promote fracture healing. To test this hypothesis, low (22 microg) and high (75 ug) doses of recombinant human PDGF-BB (rhPDGF-BB) were applied directly to femur fracture sites in BB Wistar diabetic rats that were then compared to untreated or vehicle-treated animals. rhPDGF-BB treatment significantly increased early callus cell proliferation compared to that in control specimens. Low dose rhPDGF-BB treatment significantly increased callus peak torque values (p < 0.05) at 8 weeks after fracture as compared to controls. High dose rhPDGF-BB treatment increased callus bone area at 12 weeks postfracture. These data indicate that rhPDGF-BB treatment ameliorates the effects of diabetes on fracture healing by promoting early cellular proliferation that ultimately leads to more bone formation. Local application of rhPDGF-BB may be a new therapeutic approach to treat diabetes-impaired fracture healing.

MicroCT analysis of hydroxyapatite bone repair scaffolds created via three-dimensional printing for evaluating the effects of scaffold architecture on bone ingrowth.

Recent studies have shown that it is now possible to construct tissue-engineered bone repair scaffolds with tight pore size distributions and controlled geometries using 3-D Printing techniques (3DP). This study evaluated two hydroxyapatite (HA) 8-mm diameter discs with controlled architectures in a rabbit trephine defect at 8 and 16 weeks using a 2 x 2 factorial design. Input parameters were time and scaffold void volume at two levels. Three output variables were extracted from MicroCT data: bone volume ingrowth with respect to total region of interest, bone volume ingrowth with respect to available ingrowth volume, and soft tissue volume. The experiment measured two groups--Group 1: 500-microm x 500-microm channels parallel to the scaffold's long axis and penetrating up 3-mm from the bottom. Group 2: 800-microm x 800-microm struts spaced 500 microm apart set perpendicularly to each other in each printed layer. Rendered 3-dimensional MicroCT scans and undecalcified histological slides of implants revealed good integration with the surrounding tissue, and a sizeable amount of bone ingrowth into the device. Factorial analysis revealed that the effects of time were the greatest determinant of soft tissue ingrowth, while time and its interaction with void volume were the greatest determinants of bone volume ingrowth with respect to both total and available volume.