The influence of synovial inflammation and hyperplasia on symptomatic outcomes up to 2 years post-operatively in patients undergoing partial meniscectomy.

OBJECTIVE: Synovitis is associated with pain and other symptoms in patients with knee osteoarthritis (OA), and in patients with meniscal tears even in the absence of radiographic OA. Patients undergoing arthroscopic partial meniscectomy were followed for 2 years to determine whether synovitis predicts post-operative symptoms. DESIGN: Thirty-three patients scheduled for arthroscopy were recruited for this pilot study. Symptoms were assessed using a knee pain scale, the Lysholm score, and the short form-12 (SF-12(®)) pre-operatively and at 16 weeks, 1 year and 2 years post-operatively. Synovial inflammation and hyperplasia were graded on surgical biopsies. Linear mixed effects models were tested to determine whether inflammation or hyperplasia is associated with outcome scores over time. RESULTS: Lysholm scores and SF-12(®) physical component sub-scores were worse pre-operatively in patients with inflammation (Lysholm: 52.42 [95% confidence interval (CI) 42.37, 62.47] vs 72.38 [66.03, 78.72], P < 0.001; SF-12: 36.81 [28.26, 45.37] vs 46.23 [40.14, 52.32], P < 0.05). Up to 2-years post-operatively, patients with inflammation achieved mean scores similar to those without inflammation. As a result, the mean improvement in Lysholm scores was 13.01 [1.48-24.53] points higher than patients without inflammation, P = 0.03. 33% (4/12) of patients with inflammation still had fair to poor Lysholm scores 2 years after surgery compared to 7% (1/15, P=0.14) without inflammation. No association between hyperplasia and symptoms was noted. CONCLUSIONS: In this pilot study of patients undergoing partial meniscectomy, synovial inflammation was associated with worse pre-operative symptoms, but not with poorer outcomes in the first 2 years post-arthroscopy. Larger cohorts and longer follow-up should be pursued to confirm this relationship, and determine if the initial response is sustained.

Comparable outcomes in fracture reduction and bone properties with RANKL inhibition and alendronate treatment in a mouse model of osteogenesis imperfecta.

UNLABELLED: We report a direct comparison of receptor activator of nuclear factor kappa B ligand (RANKL) inhibition (RANK-Fc) with bisphosphonate treatment (alendronate, ALN) from infancy through early adulthood in a mouse model of osteogenesis imperfecta. Both ALN and RANK-Fc decreased fracture incidence to the same degree with increases in metaphyseal bone volume via increased number of thinner trabeculae. INTRODUCTION: The potential therapeutic benefit of RANKL inhibitors in osteogenesis imperfecta (OI) is under investigation. We report a direct comparison of RANKL inhibition (RANK-Fc) with bisphosphonate treatment (ALN) from infancy through early adulthood in a model of OI, the oim/oim mouse. METHODS: Two-week-old oim/oim, oim/+, and wildtype (+/+) mice were treated with RANK-Fc 1.5 mg/kg twice per week, ALN 0.21 mg/kg/week or saline (n = 12-20 per group) for 12 weeks. RESULTS: ALN and RANK-Fc both decreased fracture incidence (9.0 ± 3.0 saline 4.4 ± 2.7 ALN, 4.3 ± 3.0 RANK-Fc fractures per mouse). Serum TRACP-5b activity decreased to 65% after 1 month in all treated mice, but increased sacrifice with RANK-Fc to 130-200% at sacrifice. Metaphyseal density was significantly increased with ALN in +/+ and oim/oim mice (p < 0.05) and tended to increase with RANK-Fc in +/+ mice. No changes in oim/oim femur biomechanical parameters occurred with treatment. Both ALN and RANK-Fc significantly increased trabecular number (3.73 ± 0.77 1/mm for oim/oim saline vs 7.93 ± 0.67 ALN and 7.34 ± 1.38 RANK-Fc) and decreased trabecular thickness (0.045 mm ± 0.003 for oim/oim saline vs 0.034 ± 0.003 ALN and 0.032 ± 0.002 RANK-Fc) and separation in all genotypes (0.28 ± 0.08 mm for oim/oim saline vs 0.12 ± 0.010 ALN and 13 ± 0.03 RANK-Fc)., with significant increase in bone volume fraction (BVF) with ALN, and a trend towards increased BVF in RANK-Fc. CONCLUSION: Treatment of oim/oim mice with either a bisphosphonate or a RANK-Fc causes similar decreases in fracture incidence with increases in metaphyseal bone volume via increased number of thinner trabeculae.

In vivo evaluation of autologous cartilage fragment-loaded scaffolds implanted into equine articular defects and compared with autologous chondrocyte implantation.

BACKGROUND: Current autologous chondrocyte implantation (ACI) techniques require 2 surgical procedures: 1 for cell harvest and 1 for reimplantation of cultured cells. A 1-step procedure is more desirable. PURPOSE: A 1-step surgical procedure using autologous cartilage fragments on a polydioxanone scaffold, or CAIS (cartilage autograft implantation system), in a clinically relevant defect (15-mm diameter) within equine femoral trochlea was compared with a 2-step ACI technique as well as with empty defects and defects with polydioxanone foam scaffolds alone. STUDY DESIGN: Controlled laboratory study. METHODS: Ten skeletally mature horses were used. Articular cartilage from the lateral trochlea of the femur was harvested arthroscopically (n = 5), and chondrocytes were cultured on small intestinal submucosa to produce ACI constructs. The CAIS procedure had cartilage harvested during defect creation to prepare minced cartilage on polydioxanone-reinforced foam. The ACI and CAIS constructs were placed in defects using polydioxanone/polyglycolic acid staples. Defects were examined arthroscopically at 4, 8, and 12 months and with gross, histological, and immunohistochemical examination at 12 months. RESULTS: Arthroscopic, histologic, and immunohistochemistry results show superiority of both implantation techniques (ACI and CAIS) compared with empty defects and defects with polydioxanone foam alone, with CAIS having the highest score. CONCLUSION: This is the first demonstration of long-term healing with strenuous exercise using ACI and CAIS in a critically sized defect. CLINICAL RELEVANCE: Given these results with the CAIS procedure, testing in human patients is the next logical step (a phase 1 human clinical study has proceeded from this work).

Local cytokine profiles in knee osteoarthritis: elevated synovial fluid interleukin-15 differentiates early from end-stage disease.

OBJECTIVE: Much of what is known about the inflammatory response in the synovial membrane (SM) of patients with osteoarthritis (OA) comes from studies of synovial tissues from end-stage disease. In this study, we sought to better characterize the inflammatory infiltrate in symptomatic patients with early signs of knee OA, and to determine how inflammatory cell populations relate to the pattern of cytokine and degradative enzyme production. METHODS: Study populations comprised patients with degenerative meniscal tears and early cartilage thinning undergoing arthroscopic procedures (early OA) and patients undergoing total knee replacement for end-stage OA. Quantitative real-time polymerase chain reaction (PCR) was used to measure expression of SM cytokines and enzymes implicated in the pathogenesis of inflammatory arthritis and OA, as well as cell lineage-specific markers. We quantified synovial fluid (SF) cytokines and enzymes by enzyme-linked immunosorbent assay (ELISA) and SM cell populations by immunohistochemistry. RESULTS: We found increased levels of SF interleukin-15 (IL-15) protein in the early knee OA patients when compared to end-stage OA. Both SF IL-15 protein and numbers of CD8 cells within SM correlated with matrix metalloproteinase-1 (MMP-1) and three levels. TNF-alpha, IL-6 and IL-21 were also detectable in the SF of the majority of patients, and IL-15 levels were associated with IL-6 levels. CONCLUSION: IL-15 is elevated in early knee OA, suggesting activation of an innate immune response in the SM. The association of IL-15 expression with CD8 transcripts and MMPs implicates this cytokine in OA pathogenesis and as a candidate therapeutic target.

Evaluation of autologous chondrocyte transplantation via a collagen membrane in equine articular defects: results at 12 and 18 months.

OBJECTIVE: To evaluate a technique of autologous chondrocyte implantation (ACI) similar to the other techniques using cell-seeded resorbable collagen membranes in large articular defects. METHODS: Autologous cartilage was harvested arthroscopically from the lateral trochlear ridge of the femur in fifteen 3-year-old horses. After culture and expansion of chondrocytes the newly created ACI construct (autologous chondrocytes cultured expanded, seeded on a collagen membrane, porcine small intestine submucosa) was implanted into 15mm defects on the medial trochlear ridge of the femur in the opposite femoropatellar joint. Using two defects in each horse, the ACI technique was compared to collagen membrane alone (CMA) and empty cartilage defects (ECDs). RESULTS: Arthroscopic evaluations at 4, 8, 12 and 18 months demonstrated that CMA was significantly worse compared to ACI or ECD treatments, with ACI having the best overall subjective grade. Overall raw histological scores demonstrated a significant improvement with ACI compared to either CMA or ECD treated defects and ACI defects had significantly more immunohistochemical staining for aggrecan than CMA or ECD treated defects (with significantly more type II collagen in ACI and ECD compared to CMA defects) at 12 and 18 months. CONCLUSIONS: Histologic and immunohistochemistry results from this long-term randomized study are particularly encouraging and demonstrate superiority with the ACI technique. Although there is no comparable study published with the traditional ACI technique in the horse (or with such a large defect size in another animal model), the use of a solid autologous cell-seeded-constructed implant would appear to offer considerable clinical advantages.

Elevated high-sensitivity C-reactive protein levels are associated with local inflammatory findings in patients with osteoarthritis.

OBJECTIVE: C-reactive protein (CRP) has been associated with disease progression in patients with osteoarthritis (OA), but the reasons for this remain unclear. We hypothesized that higher CRP would be related to local inflammatory findings in the joints of patients with OA. METHODS: Plasma and synovial membrane specimens from 54 OA patients undergoing total hip or knee arthroplasty or arthroscopy were obtained. Synovial fluid was obtained from 25 of these patients. Hematoxylin and eosin stained synovial membrane sections were scored for degree of inflammatory cell infiltration. Plasma high-sensitivity CRP (hsCRP) levels, and serum and synovial fluid interleukin (IL)-6 and IL-1beta levels were measured by enzyme-linked immunosorbent assay. RESULTS: Fifty-seven percent of patients with idiopathic OA had inflammatory infiltrates within the synovial membrane. The mean hsCRP level in patients with inflammatory infiltrates was significantly higher than those without inflammation (4.7 +/- 5.0 mg/L vs 1.7 +/- 3.6 mg/L, P = 0.003). There were significant correlations between hsCRP levels and synovial fluid IL-6 (r = 0.64, P = 0.0006), degree of synovial inflammatory infiltration (r = 0.43, P = 0.002), and body mass index (r = 0.31, P = 0.02). Multivariate analysis indicated that only degree of inflammatory infiltrate was significantly associated with hsCRP level (P = 0.026). CONCLUSION: These results suggest that systemic hsCRP levels reflect synovial inflammation in OA patients, perhaps by means of synovial IL-6 production. Future studies are needed to clarify how these infiltrates and their products may contribute to disease pathogenesis.

Comparison of mineral quality and quantity in iliac crest biopsies from high- and low-turnover osteoporosis: an FT-IR microspectroscopic investigation.

Fourier-transform infrared microspectroscopy (FTIRM) allows analysis of mineral content, mineral crystal maturity and mineral composition at approximately 10-micron spatial resolution. Previous FTIRM analyses comparing 4-micron thick sections from non-decalcified iliac crest biopsies from women with post-menopausal osteoporosis, as contrasted with iliac crest tissue from individuals without evidence of metabolic bone disease, demonstrated significant differences in average mineral content (decreased in osteoporosis) and mineral crystal size/perfection (increased in osteoporosis). More importantly, these parameters, which vary throughout the tissue in relation to the tissue age in healthy bone, showed no such variation in bone biopsies from patients with osteoporosis. The present study compares the spatial and temporal variation in mineral quantity and properties in trabecular bone in high- and low-turnover osteoporosis. Specifically, six biopsies from women (n=5) and one man with high-turnover osteoporosis (age range 39-77) and four women and two men with low turnover osteoporosis (age range 37-63) were compared to ten "normal" biopsies from three men and seven woman (age range: 27-69). "High turnover" was defined as the presence of increased resorptive surface, higher than normal numbers of osteoclasts and greater than or equal to normal osteoblastic activity. "Low turnover" was defined as lower than normal resorptive surface, decreased osteoclast number and less than normal osteoblastic activity. Comparing variations in FTIR-derived values for each of the parameters measured at the surfaces of the trabecular bone to the maximum value observed in multiple trabeculae from each person, the high-turnover samples showed little change in the mineral: matrix ratio, carbonate: amide I ratio, crystallinity and acid phosphate content. The low-turnover samples also showed little change in these parameters, but in contrast to the high-turnover samples, the low-turnover samples showed a slight increase in these parameters, indicative of retarded, but existent resorption and formation. These data indicate that FTIR microspectroscopy can provide quantitative information on mineral changes in osteoporosis that are consistent with proposed mechanisms of bone loss.

Distribution of collagen cross-links in normal human trabecular bone.

UNLABELLED: Infrared imaging analysis of normal human iliac crest biopsy specimens shows a characteristic spatial variation in the nonreducible:reducible collagen cross-links at trabecular surfaces, depending on the surfaces' metabolic status. INTRODUCTION: Bone is a composite material consisting of mineral, collagen, non-collagenous proteins, and lipids. Bone collagen, mainly type I, provides the scaffold on which mineral is deposited and imparts specific mechanical properties, determined in part by the amount of collagen present, its orientation and fibril diameter, and the distribution of its cross-links. MATERIALS AND METHODS: In this study, the technique of Fourier transform infrared imaging (FTIRI) was used to determine the ratio of nonreducible:reducible cross-links, in 2- to 4-microm-thick sections from human iliac crest biopsy specimens (N = 14) at trabecular surfaces as a function of surface activity (forming versus resorbing), with an approximately 6.3-mm spatial resolution. The biopsy specimens were obtained from patients devoid of any metabolic bone disease based on histomorphometric and bone densitometric parameters. RESULTS AND CONCLUSIONS: Distributions of collagen cross-links within the first 50 mm at forming trabecular surfaces demonstrated a progressive increase in the nonreducible:reducible collagen cross-link ratio, unlike in the case of resorbing surfaces, in which the collagen cross-links ratio (as defined for the purposes of the present report) was relatively constant.

The anatomy and histology of the rotator interval capsule of the shoulder.

Forty-seven rotator interval regions from fetuses and 10 fresh-frozen rotator interval regions from adult cadavers were evaluated by gross dissection and light microscopy. Specimens from adults also were evaluated with ultrasound and magnetic resonance imaging. An analysis of 37 fetal specimens (> 14 weeks gestation) revealed two rotator interval types: Type I (9 of 37) was defined by a contiguous bridge of capsule consisting of poorly organized collagen fibers. A Type II rotator interval (28 of 37) had a complete defect covered by only a thin layer of synovium. Similar to the Type II rotator interval in the fetus, a rotator interval defect was present in six of eight specimens from adults. Histologically, the capsular tissue within the rotator interval consisted of poorly organized collagen fibers in specimens from the fetus and adult. Maximal opening of the rotator interval was seen by ultrasound with internal rotation and downward traction of the hyperextended arm in the coronal, oblique, and sagittal planes. Magnetic resonance imaging of the rotator interval region permitted anatomic evaluation. The complete absence of tissue in 28 of 37 fetuses suggests that the rotator interval defect is congenital. The authors recommend that surgeons carefully evaluate the integrity of the tissue within the rotator interval. When rotator interval closure is desired such as in patients with a persistent sulcus sign after arthroscopic stabilization, suturing the edges of more substantial tissue immediately adjacent to the boundaries of the rotator interval region would seem prudent.

An unexpected outcome during testing of commercially available demineralized bone graft materials: how safe are the nonallograft components?

STUDY DESIGN: Radiographic and histologic analyses of commercially available bone graft materials were performed. OBJECTIVE: To compare the osteoinductive efficacy of commercially available demineralized bone matrix material. SUMMARY OF BACKGROUND DATA: The relative in vivo bone formation and toxicology of the nonallograft components the make up various commercially available demineralized bone matrix products are not known. METHODS: An in vivo bone formation model was used in 30 athymic rats. Six different bone grafting materials were tested in subcutaneous and intermuscular locations. After 4 weeks, radiographic and histologic testing of bone formation was performed. RESULTS: Eight of nine rats implanted with Grafton demineralized bone matrix products died 1 to 4 days after implantation of the bone graft material. None of the remaining 10 animals implanted with the four other grafting materials died. The experiment was modified and completed with a lower dose of bone graft material. Pathologic analysis indicated that the cause of death was hemorrhagic necrosis of the kidneys, most likely caused by a toxic effect on the glomeruli and tubules. A possible causative factor may have been the glycerol in the graft material. CONCLUSIONS: Although the volume of Grafton product per kilogram of body weight used in this study was approximately eight times the maximum volume used in humans, the authors believe that this data must be reported because this product is used substantially in clinical settings. In addition, the osteoinductive performance and relative safety of the nonallograft components in all commercially available demineralized bone grafts are not known.

Spinal epidural hemangioma related to pregnancy.

We report the case of a 39-year-old woman with adolescent idiopathic scoliosis presenting with myelopathy secondary to a spinal epidural hemangioma. MRI showed an epidural soft tissue mass within the spinal canal between T5 and T9 with severe spinal cord compression. Symptoms had a temporal relationship to her pregnancy. Surgical removal of the epidural hemangioma rapidly relieved her symptoms and neurologic deficits. Follow-up examination 2 years later demonstrated normal motor and sensory function, without any neurologic sequelae or progression of deformity.

The developmental anatomy of the neonatal glenohumeral joint.

The embryologic development of the capsular ligaments, synovial lining, rotator cuff, and bony structures of the shoulder is incompletely understood. The purpose of this study is to report the gross and microscopic anatomy of the developing glenohumeral joint on the basis of dissections of fetal shoulder specimens. After Institutional Review Board approval from our hospital, 51 shoulders in 37 fetal specimens were obtained from cases of fetal demise. The gestation time of these specimens ranged from 9 to 40 weeks. The morphology of the capsule, labrum, and associated ligaments were studied by dissection under a dissecting microscope. High-resolution radiographs were made, and sections were processed for routine histology. There was noted to be minimal variation in the shape and slope of the acromion. The coracoid was much larger in relation to the shoulder than in the mature shoulder. The coracoacromial ligament was grossly evident at this stage of development, with distinct anterolateral and posteromedial bands in this ligament. The inferior glenohumeral ligament was seen as a prominent thickening in the capsule, whereas the middle and superior glenohumeral ligaments were thinner and more difficult to identify as distinct structures. Upon histologic examination, the inferior glenohumeral ligament was seen to consist of several layers of organized collagen fibers. The inferior glenohumeral ligament inserted into the labrum and margin of the glenoid. The capsule was much thinner in the region superior to the inferior glenohumeral ligament. A rotator interval capsular defect was often present, and the coracohumeral ligament was seen as a distinct structure as early as 15 weeks. A bare spot in the glenoid was not observed. This study indicates that some of the important functional elements of the structure of the mature human shoulder are present early in development, including the glenohumeral and coracohumeral ligaments. The coracoacromial ligament plays a significant role in the formation of the coracoacromial arch in the neonatal shoulder. The presence of a capsular rotator interval indicates that this aspect of capsular anatomy is congenital.

The prevalence and clinicopathological appearance of extension of osteonecrosis in the femoral head.

In about 50% of cases, osteonecrosis of the femoral head is known to occupy more than one site. There is controversy as to whether a single focus may increase in size. We have reviewed 606 consecutive femoral heads which had been surgically removed for osteonecrosis. Extension of osteonecrosis was observed in only two (0.3%) and was confirmed histopathologically by the enlargement of the necrotic segment beyond the repair zone formed for the primary necrosis into the adjacent, previously uninvolved bone. In both cases, the necrotic regions were wedge-shaped and occupied over 80% of the femoral head. It appears that an increase in size is extremely rare and that osteonecrosis is due to a single event. Our findings may be of value in assessing the use of joint-salvage procedures for osteonecrosis of the femoral head.

Osteonecrosis in the rheumatoid femoral head.

OBJECTIVE: To define the prevalence and pathological spectrum of femoral head osteonecrosis in patients with rheumatoid arthritis (RA) and to correlate its presence with disease related clinical and therapeutic factors. METHODS: A total of 545 primary total hip arthroplasties performed in 507 patients with RA were identified. A historical review of each patient's rheumatoid disease and treatment as well as pathological review of each femoral head specimen was performed. RESULTS: Osteonecrosis was identified in 66 specimens (12.1%) in one of 2 discrete forms. Thirty-two specimens (5.9%) contained classic subchondral avascular necrosis. Thirty-four specimens (6.2%) contained osteonecrosis in association with degenerative changes (within regions of sclerotic and eburnated subchondral bone), but not classic avascular necrosis. Remaining femoral head specimens were characterized by inflammatory arthritis (431 specimens) or degenerative joint disease (48 specimens). Corticosteroid therapy was used in 81% of patients with avascular necrosis and 68% with degenerative osteonecrosis. This was significantly greater prevalence than in patients without osteonecrosis (33%). Average daily prednisone dosage was 8 mg and no association between dosage and the presence of osteonecrosis was identified. No correlation between pathological findings and clinical disease severity was identified. In 5 of 27 specimens showing classic avascular necrosis and 11 of 34 containing degenerative osteonecrosis, no steroid treatment had been administered. CONCLUSION: Femoral head osteonecrosis is present in about 12% of patients with RA at hip arthroplasty, and occurs in 2 forms -- classic avascular necrosis and degenerative necrosis. Both forms are significantly associated with corticosteroid use. "Low dose" therapy does not protect patients against the development of osteonecrosis. Additionally, baseline prevalence of osteonecrosis of about 3% occurs in the absence of steroid use and may be related to the underlying inflammatory diseases. Despite its association with osteonecrosis the net effect of corticosteroid therapy on the natural history of rheumatoid hip disease remains unclear.

Clinical and pathologic findings in hemochromatosis hip arthropathy.

During a 9-year period, 15 patients with hemochromatosis hip arthropathy required 19 total hip arthroplasties for disabling hip pain. Preoperative presentation, hip function, pathologic evaluation of the femoral head, and radiographic findings were reviewed. Postoperative followup averaging 5.7 years (range, 2-11 years) was performed to assess hip pain and function after total hip arthroplasty. The average preoperative Hospital for Special Surgery hip score was 15 points (range, 4-24 points), and this improved to 30 points (range, 4-38 points) after total hip arthroplasty. Only one of 15 patients required revision surgery at 10 years for acetabular loosening. All other patients were pain free, with improved function at latest followup. Histologic evaluation of the resected femoral heads revealed evidence of primary or secondary osteonecrosis in seven of 19 (37%) specimens. Articular cartilage avulsion at the level of the tidemark was identified in eight of 19 (42%) specimens, and calcium pyrophosphate deposition was identified in five of 19 (26%) specimens. These pathologic findings suggest a predictable progression of the arthritic process in patients with hemochromatosis.

Chronic ankle pain caused by osteoid osteoma of the neck of the talus.

Five patients with osteoid osteomas of the talar neck were treated at the Hospital for Special Surgery between 1981 and 1992. The course of care leading to definitive diagnosis and treatment was reviewed. All five of the patients had night pain relieved by aspirin or nonsteroidal anti-inflammatory drugs. One of the five reported associated trauma. The average time from onset of symptoms to correct diagnosis was 2.5 years. Juxta-articular osteoid osteoma can cause a small spur that resembles a traction spur on the neck of the talus. Anterior ankle impingement was the most common misdiagnosis. Initial treatments included arthroscopic spur debridement or synovectomy, casting for fracture, and repeated nerve blocks for reflex sympathetic dystrophy. The five patients were cured by en bloc excision of the lesion. In the diagnosis of osteoid osteoma, a history of relief of pain with aspirin is important. Plain radiographs and a bone scan are useful. Fine cut computed tomography scanning or magnetic resonance imaging are the best studies for making a definitive diagnosis. Localization by computed tomography guided needle placement or intraoperative radionuclide scanning are recommended to find the lesion for excision. Intraoperative radiographs of the excised lesion should be used to confirm complete removal.

FTIR microspectroscopic analysis of normal human cortical and trabecular bone.

Fourier transform infrared microspectroscopy (FTIRM) has been used to study the changes in mineral and matrix content and composition in replicate biopsies of nonosteoporotic human cortical and trabecular bone. Changes in osteonal bone in these same samples were reported previously. Spectral maps along and across the lamellae were obtained from iliac crest biopsies of two necropsy cases. Mineral:matrix ratios, calculated from the integrated areas of the phosphate nu1, nu3 band at 900-1200 cm-1 and the amide I band at approximately 1585-1725 cm-1, respectively, were relatively constant in both directions of analysis, i.e., along and across the lamellae. Analysis of the components of the nu1, nu3 phosphate band with a combination of second-derivative spectroscopy and curve fitting revealed the presence of 11 major underlying moieties. Of these, the ratio of the relative areas of the two underlying bands at approximately 1020 and approximately 1030 cm-1 has been shown to be a sensitive index of variation in crystal perfection in both human osteonal bone and in synthetic, poorly crystalline apatites. This ratio was calculated in both cortical and trabecular bone from human iliac crest biopsies along and across the lamellae. The ratio decreased, going from the periosteum to the medullary cavity in the cortical bone, and from the periphery towards the center of trabeculae. These observations were consistent within serial sections obtained from the same biopsy, multiple biopsies obtained from the same necropsy specimen, and biopsies obtained from the two different necropsy specimens. The results presented here along with previously reported changes in osteonal bone show a relation between bone age and "crystallinity/maturity" (a parameter dependent on crystallite size, hydroxyapatite-like stoichiometry, abundance of substituting ions such as CO32-; the more crystalline/mature, the more hydroxyapatite-like stoichiometry, the bigger the crystallite size, the less the ion substitution by ions such as CO32-) as deduced by the 1020/1030 cm-1 ratio. Invariably, younger normal bone is less mature/crystalline than older. These results provide a "baseline" for description of mineral properties, to which diseased bones may be compared.

FTIR microspectroscopic analysis of human iliac crest biopsies from untreated osteoporotic bone.

Historically, osteoporosis has been defined as a disease in which there is "too little bone, but what there is, is normal." As a result of research design and sample selection limitations, published data contradict and confirm the historical definition. Because of these limitations, it has been hard to assess the contribution of mineral quality to mechanical properties, and to select therapeutic protocols that optimize bone mineral properties. The coupling of an optical microscope to an infrared spectrometer enables the acquisition of spectral data at known sites in a histologic section of mineralized tissue without loss of topography and/or orientation. The use of second-derivative spectroscopy coupled with curve-fitting techniques allows the qualitative and quantitative assessment of mineral quality (crystallite size and perfection, mineral:matrix ratio) at well-defined morphologic locations. We have previously applied these techniques to the study of normal human osteonal, cortical, and trabecular bone. The results indicated that the newly deposited bone mineral is less "crystalline/mature" than the older one. In the present study, Fourier transform infrared microspectroscopy (FTIRM) was applied to the study of human osteonal and cortical bone from iliac crest biopsies of untreated osteoporotic patients. The hypothesis tested was that osteoporotic bone mineral is monotonically different in its properties expressed as "crystallinity/maturity" than the normal. The results indicate significant differences in the mineral properties as expressed by crystal size and perfection, with the mineral from osteoporotic bone being more crystalline/mature than the normal.

FTIR microspectroscopic analysis of human osteonal bone.

Fourier Transform Infrared Microspectroscopy (FTIRM) has been used to study the changes in mineral and matrix content and composition in replicate biopsies of nonosteoporotic human osteonal bone. Spectral maps in four orthogonal directions (in 10 microm steps) from the centers towards the peripheries of individual osteons were obtained from iliac crest biopsies of two necropsy cases. Mineral to matrix ratios, calculated from the ratio of integrated areas of the phosphate nu1,nu3 band at 900-1200 cm-1 to the amide I band at 1585-1725 cm-1, increased from the center to the periphery of the osteon. The total carbonate (based on the nu2 band at approximately 850-900 cm-1) to phosphate nu1,nu3 ratio decreased as the mineral to matrix ratio increased. Analysis of the nu2 CO32- band with a combination of second-derivative spectroscopy and curve fitting revealed a decrease in "labile" carbonate, a slight decrease in Type A and a slight increase in Type B carbonate from the center to the periphery of the osteon. Similar analysis of the components of the nu1,nu3 phosphate band with a combination of second-derivative spectroscopy and curve fitting revealed the presence of 11 major underlying moieties. These components were assigned by comparison with published frequencies for apatite and acid-phosphate containing calcium phosphates. The most consistent variations were alterations in the relative percent areas of bands at approximately 1020 and approximately 1030 cm-1, which had previously been assigned to nonstoichiometric and stoichiometric apatites, respectively. This ratio was used as an index of variation in crystal perfection throughout the osteon. This ratio decreased as the mineral to matrix ratio increased. The reproducibility of these parameters at multiple sites in multiple biopsies suggests their applicability for the analysis of mineral changes in disease.