Postmenopausal women with osteoarthritis and osteoporosis show different ultrastructural characteristics of trabecular bone of the femoral head
© Shen et al; licensee BioMed Central Ltd. 2009
Received: 22 October 2008
Accepted: 09 April 2009
Published: 09 April 2009
Osteoporosis (OP) and osteoarthritis (OA) are public health diseases affecting the quality of life of the elderly, and bring about a heavy burden to the society and family of patients. It has been debated whether or not there is an inverse relationship between these two disorders.
To compare the exact difference in bone tissue structure between osteoporosis and osteoarthritis, we observed the ultrastructure of trabecular bone from the femoral heads using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). A total of 15 femoral head specimens from postmenopausal women were collected during the procedures of total or hemi hip replacement (OP, n = 8; OA, n = 7). The morphologic structure of the trabecular bone, collagen fibers, resorption lacuna and osteoblasts were observed.
Under SEM, osteoporotic trabeculae appeared to be thinning, tapering, breaking and perforating. A number of resorption lacunae of various shapes were seen on the surface of the trabeculum. The collagen fibers of lacuna were resorbed. On occasion, naked granular bone crystals could be found. In the OA group, the trabecular bone looked thick with integrated structure. Reticular and granular new bone could be found. The trabeculum was covered by well-arranged collagen fibers around the resorption lacuna. In the OP group, under TEM, marginal collagen fibers were observed to be aligned loosely with enlarged spaces. A few inactive osteoblasts and no inflammatory cells were seen. In the OA group, the collagen fibers inside the trabeculum were arranged in a dense manner with many active osteoblasts and inflammatory cells infiltrating the matrix.
We found significant differences in the trabecular bone, collagen fibers, lacunae and osteoblasts between postmenopausal women with OP and OA. These findings support the hypothesis that there is an inverse relationship between OP and OA.
Osteoporosis (OP) and osteoarthritis (OA) are two common diseases that severely influence quality of life, especially for the elderly. OP, characterized by the reduction in the amount of bone and deterioration of bone microarchitecture, is considered to be the consequence of an imbalance between bone formation and resorption. It also makes bone susceptible to fracture with increased bone fragility. OP is clinically defined as a condition in which bone mineral density (BMD) is at least 2.5 standard deviations (also referred to as 'T-score') below the mean of the young adult population by World Health Organization (WHO) . OA is manifested by progressive degeneration of articular cartilage which is believed to be usually caused by articular cartilage erosion and chondrocyte damage . The subchondral bone may play an important role in the pathogenesis of OA . The sclerotic subchondral bone is considered to weaken the articular cartilage by impairing its ability to absorb mechanical shock, thereby influencing the progression of OA . Although both OA and OP are strongly related to age and metabolism, they are multifaceted conditions influenced by mechanical and genetic factors [5–10].
Although the relationship between OA and OP remains controversial, an inverse relationship has been more widely accepted [11–14]. In clinical setting, both diseases rarely occurred in the same patient . The femoral heads from osteoporotic fractures were found well preserved in earlier studies. Comparison of bone mineral densities (BMDs) in OA, OP and normal controls, the BMDs of osteoarthritic patients were the highest [16–19]. Even if patients with OA do suffer from osteoporotic fractures, the age at fracture occurrence is usually much older, which indicates that OA might have a protective effect on fracture .
The trabeculae in patients with OP have lower strength and are of poorer quality , whereas sclerotic subchondral trabecular bone is found in those with OA. However, the increase of stiffness in OA does not mean higher strength. Ding et al  reported that the thickness of trabeculae in early-staged OA patients increased significantly, but the strength of the subchondral trabecular bone was still weaker than healthy controls. Although the relationship between OA and OP has been investigated with regard to subchondral bone plates  or composition and mechanical properties of cancellous bone , the ultrastructure of trabecular bone has not been compared between these two diseases using both scanning electron microscopy (SEM) and transmission electron microscopy (TEM). An exploration of the ultrastructure of the trabecular bone, which contributes to the mechanical features, might be helpful to explain the real relationship between OA and OP populations. The aim of this study was to to investigate the trabecular ultrastructure between OA and OP using SEM and TEM with a working hypothesis for an inverse relationship between OA and OP.
Femoral heads were obtained from 15 postmenopausal women with an average age of 76.8 (63–86 yrs) during hip replacement operations. Eight patients diagnosed with osteoporotic femoral neck fracture undertook a hemi-hip replacement, while the other seven with primary osteoarthritis sustained a total hip arthroplasty. To avoid disturbance of age and hormone level, each donor has at least five years menopause history. Each sample of articular cartilage from the OA group had severe erosion as defined by Outerbridge grade IV . To ensure more consistent bone quality, patients with old osteoporotic fracture were precluded from this series. Any patients with osteomalacia, multiple myeloma, rheumatoid arthritis, or secondary osteoporosis due to hormone therapy were excluded from the OP group. Likewise, patients with congenital or acquired hip dysplasia, gout, rheumatoid arthritis or avascular necrosis of the femoral head were excluded from the OA group. The investigation was approved by institutional review board of our institution. In accordance with local ethical standards, informed consent was obtained from patients.
The femoral head was bivalved in the coronal plane with a sharp osteotome. The exposed surface was rinsed with saline solution repeatedly to remove blood and bone marrow. Then specimens, 5 mm × 5 mm × 5 mm in size for SEM and 2 mm × 2 mm × 2 mm for TEM, were harvested from the coronal medial plane from the trabecular structure of the femoral head, 1.5 cm below the joint surface [25, 26]. All specimens were fixed with 2% glutaraldehyde solution, washed with 0.1 M sodium cacodylate buffer, and post-fixed with 1% osmium tetroxide. After dehydrating with an alcohol gradient series, different protocol was performed for SEM and TEM procedures. For SEM, after dehydrating with isoamyl acetate again, the specimen was dried using a critical point dryer with HCP-2. After being coated with a layer of gold, all specimens were studied under a scanning electron microscope (QUANTA-200, Philips, Eindhoven, The Netherlands). For TEM, each specimen was doubly replaced with propylene oxide, soaked with epoxy resin, and embedded in oven of 60°C for 48 hours. Specimens were then sectioned into ultra-thin slices, dyed with citric acid lead, and examined under a transmission electron microscope (CM-120, Philips, Eindhoven, The Netherlands).
Many resorption lacunae with oval, narrow oval or spindle shapes could be seen over the icicle-like trabeculae. The margin of the lacuna showed an irregular, perforated appearance. Adjacent lacunae were observed to have coalesced and fused together. Smooth and regular collagen fibrils could be discerned among the lacunae, namely on the unresorbed surface. Under high magnification, the collagen fibrils were of uniform size, existing in a parallel array orientation and showing oblique and finer connecting fibrils. Round, oval, or residual inorganic granules with irregular shape could be observed in the lacuna. On the bottom of the lacuna, tight or loose collagen fibrils presented irregular arrangement and breakage (Fig. 2B).
Resorption of the fibrils was also observed on the surface of the trabeculae among lacunae. Under high magnification, the collagen fibrils appeared loosely scattered and uneven, whereas the lacuna was shallow and empty with a perforated margin (Fig. 2C).
Thinner collagen fibrils 0.5–1 μm in diameter and 5–10 μm in length, which represented the newly formed fibrils, emerged from inside the lacuna on the icicle-like trabecular bone (Fig. 3B). These fibrils appeared in a clear border arranged in a regular and parallel order. This origination extended from one side, filling up the bottom, towards the para-lacunar region on the opposite side (Fig. 3C).
The reticular new bone tissue formed by the thin fibril mesh could be found adhering to the surface of the trabeculae (Fig. 4C). Thicker fibers fused together to form a plate-like structure (Fig. 4D). A number of resorption lacunae generated by osteoclasts made the trabeculae a porous appearance. Thin and regular collagen fibrils were tightly arranged (Fig. 4E). A small amount of new bone formation was found in some lacunae with the occasional microfracture. Granular new bone appeared beside the reticular bone. The granular new bone gathered together with approximate size filling the space between the trabeculae (Fig. 4F). In comparison with the OP group, no inorganic bone crystal granules were present.
Bone tissue is composed of two different components: organic and inorganic, which determine the toughness and rigidity, respectively. Both of these substances also serve the mechanical strength of the bone. Bone structure is composed of cortical and trabecular bone. While the former mainly bears mechanical load, the latter is more sensitive to hormones or other biological factors that are involved in modulating bone metabolism.
The mechanical properties of the trabecular bone are influenced by its microarchitechture, such as trabecular number, thickness, and spacing. Dilworth et al  noted significant difference in mean trabecular thickness between groups fed with or without zinc supplementation diets in an electron microscope study. The ratio of the trabecular nodes to terminations was considered as one of the important factors affecting bone strength. Other authors [28, 29] also observed cortical bone under electron microscope.
Byers et al , who investigated more than 100 femoral heads excised from patients with femoral neck fracture, did not find any osteoarthritic change. In another epidemiological study in Jerusalem, the authors found that the incidence of OP and OA were 16.1% and 4.1%, respectively. However, only 0.5% had both diseases simultaneously . Li et al  reported a trabecular bone loss of 17% in osteoporotic femoral heads, while a 60% increase was observed in patients with OA. The higher bone quantity and better mechanical quality could partly explain why femoral neck fractures were so rare in those people with OA . However, Papadakis M et al  observed that the degree of lumbar lordosis was not associated with the presence of OA or OP. The reasons for the lack of difference may be due to, we believe, the size of sample, the criteria of subgroup, and the age of the patients.
In our present study, we found significant differences in the ultrastructure of the trabecular bone between OA and OP groups. Not only the structure but also collagen fibrils were shown intact in OA, whereas destructive changes were noted in OP. Meanwhile, more new bone formation could be observed in osteoarthritic donors. However, thinning and sparse trabecular bone was the most outstanding manifestation in the OP group. In addition, severe destructive changes in the rod-like trabeculae, such as sharpening and breakage, were shown in all specimens from the OP group. The similar appearance also occurred in the collagen fibrils on the trabecular surface to some extent. Any impairment in continuity and integrity might have a potential effect on bone strength. Results from TEM also supported that the increase in resorptive activity noted in OP patients might be related to more bone loss in comparison with OA. However, obviously increased new bone tissue in osteoarthritic samples implied that bone formation was more active than bone resorption. This finding is consistent with that reported by other authors . As we know, most investigations by TEM focus on the cartilage or synovium of femoral head in OA. So the information about trabecular bone in OA under TEM was considerably limited.
The superficial layer of fibrils in OP varied in diameter, while some of them vanished to some degree. Similar changes were also observed in the deep layer fibrils. On the contrary, the collagen fibrils in both superficial and deep layers in OA remained intact and regular. Although this provides better bone toughness, more new bone tissue formation also increases bone stiffness, as described by the hypothesis proposed by Dequeker. 
Different kinds of new bone formation were observed in both OA and OP groups in this study. New collagen fibrils and bone matrix in the lacunae rarely existed in patients with OP, whereas reticular and granular new bone was shown widely in those with OA. Regularly arranged new collagen fibrils in the resorption lacunae implied that more active bone formation dominated bone turnover in osteoarthritic patients. However, those resorption lacunae also indicated high level of resorption in the OA group.
Collagen fibrils and matrix components could be synthesized and secreted by osteoblasts. In the present study, significant differences in appearance, number, and cellular organs of osteoblasts were shown between OA and OP groups. Osteoblasts in OP group demonstrated little function and scattered sparsely. Moreover, new osteoblasts were hardly seen in these osteoporotic donors. Conversely, a great number of osteoblasts with active function were noted in all specimens with OA. Thus, we suggested that more bone mass in OA population might be due to more active bone formation.
This study was limited by the nature of study as SEM and TEM is a 2-D method. By contrast, a micro-CT reconstruction is a powerful method to delineate structural characteristics of the trabecular bone in a 3-D fashion . Therefore, a comparison between the OP and OA groups using 3-D micro-CT reconstruction would be necessary in the further studies.
Another limitation of this study might be the lack of quantitative data from electron microscopic findings. In fact, however, a 2-D histomorphometric analysis of cartilage and subchondral bone in postmenopausal women with OA and OP was made in our previous study . In that study, lower bone volume fraction (BV/TV), trabecular thickness (Tb.Th), trabecular number (Tb.N) and the ratio of nodes to termini (Nd/Tm) were demonstrated in OP patients than OA patients, whereas increased trabecular space (Tb.Sp) was noted in these OP patients.
In summary, we found significant differences in the ultrastructure of the trabecular bone between postmenopausal women with osteoporosis and osteoarthritis using SEM and TEM. These findings not only suggest totally different mechanism and progression of two common diseases, but also support the hypothesis that there is an inverse relationship between OA and OP. Bone resorption and formation activity of the trabecular bone prevail in OP and OA patients, respectively.
This study was supported by Science and Technology Commission of Shanghai Municipality (08JC1415800, 08411950100)
- World Health Organization Study Group: Assessment of fracture risk and its application to screening for postmenopausal osteoporosis. World Health Organ Tech Rep Ser. 1994, 843: 1-129.Google Scholar
- Brown SJ, Pollintine P, Powell DE, Davie MW, Sharp CA: Regional differences in mechanical and material properties of femoral head cancellous bone in health and osteoarthritis. Calcif Tissue Int. 2002, 71: 227-234. 10.1007/s00223-001-2102-y.View ArticlePubMedGoogle Scholar
- Radin EL, Paul IL: Does cartilage compliance reduce skeletal impact loads? The relative force-attenuating properties of articular cartilage, synovial fluid, periarticular soft tissues and bone. Arthritis Rheum. 1970, 13: 139-144. 10.1002/art.1780130206.View ArticlePubMedGoogle Scholar
- Li B, Aspden RM: Composition and mechanical properties of cancellous bone from the femoral head of patients with osteoporosis or osteoarthritis. J Bone Mine Res. 1997, 12: 641-651. 10.1359/jbmr.1922.214.171.1241.View ArticleGoogle Scholar
- Page WF, Hoaglund FT, Steinbach LS, Heath AC: Primary osteoarthritis of the hip in monozygotic and dizygotic male twins. Twin Res. 2003, 6: 147-151. 10.1375/136905203321536272.View ArticlePubMedGoogle Scholar
- Uitterlinden AG, Burger H, Huang Q, Yue F, McGuigan FE, Grant SF, Hofman A, van Leeuwen JP, Pols HA, Ralston SH: Relation of alleles of the collagen type Iα1 gene to bone density and the risk of osteoporotic fractures in postmenopausal women. N Eng J Med. 1998, 338: 1016-1021. 10.1056/NEJM199804093381502.View ArticleGoogle Scholar
- Yamada Y, Miyauchi A, Takagi Y, Nakauchi K, Miki N, Mizuno M, Harada A: Association of a polymorphism of the transforming growth factor β-1 gene with prevalent vertebral fractures in Japanese women. Am J Med. 2000, 109: 244-247. 10.1016/S0002-9343(00)00468-X.View ArticlePubMedGoogle Scholar
- Szulc P, Munoz F, Claustrat B, Garnero P, Marchand F, Duboeuf F, Delmas PD: Bioavailable estradiol may be an important determinant of osteoporosis in men: the MINOS Study. J Clin Endocrinol Metab. 2001, 86: 192-199. 10.1210/jc.86.1.192.PubMedGoogle Scholar
- Riggs BL, Kholsa S, Melton LJ: A unitary model for involutional osteoporosis: estrogen deficiency causes both type 1 and type 2 osteoporosis in postmenopausal women and contributes to bone loss in aging men. J Bone Miner Res. 1998, 13: 763-773. 10.1359/jbmr.19126.96.36.1993.View ArticlePubMedGoogle Scholar
- Falahati-Nini A, Riggs BL, Atkinson EJ, O'Fallon WM, Eastell R, Khosla S: Relative contributions of testosterone and estrogen in regulating bone resorption and formation in normal elderly men. J Clin Invest. 1998, 106: 1553-1556. 10.1172/JCI10942.View ArticleGoogle Scholar
- Dequeker J, Boonen S, Aerssens J, Westhovens R: Inverse relationship osteoarthritis-osteoporosis: what is the evidence? what are the consequences?. Br J Rheumatol. 1996, 35: 813-818. 10.1093/rheumatology/35.9.813.View ArticlePubMedGoogle Scholar
- Dai LY: The relationship between osteoarthritis and osteoporosis in the hip. J Orthop Rheumatol. 1996, 9: 214-216.Google Scholar
- Dai LY: The relationship between osteoarthritis and osteoporosis in the spine. Clin Rheumatol. 1998, 17 (1): 44-46. 10.1007/BF01450957.View ArticlePubMedGoogle Scholar
- Jiang LS, Zhang ZM, Jiang SD, Chen WH, Dai LY: Differential bone metabolism between postmenopausal women with osteoarthritis and osteoporosis. J Bone Miner Res. 2008, 23: 475-483. 10.1359/jbmr.071114.View ArticlePubMedGoogle Scholar
- Verstraeten A, Van Ermen H, Haghebaert G, Nijs J, Geusens P, Dequeker J: Osteoarthrosis retards the development of osteoporosis. Observation of the coexistence of osteoarthrosis and osteoporosis. Clin Orthop. 1991, 264: 169-177.PubMedGoogle Scholar
- Urist MR: Observations bearing on the problem of osteoporosis. Bone as a tissue. Edited by: Bodahl K. 1960, New York: McGraw-Hill, 18-23.Google Scholar
- Smith RW, Rizek J: Epidemiologic studies of osteoporosis in women of Puerto Rico and Southeastern Michigan with special reference to age, race, national origin and to other related or associated findings. Clin Orthop. 1966, 45: 31-48.View ArticlePubMedGoogle Scholar
- Foss MVL, Byers PD: Bone density, osteoarthrosis of the hip and fracture of the upper end of the femur. Ann Rheum Dis. 1972, 31: 259-264. 10.1136/ard.31.4.259.View ArticlePubMedPubMed CentralGoogle Scholar
- Dequeker J, Burssens A, Creytens G, Bouillon R: Ageing of bone: its relation to osteoporosis and osteoarthrosis in postmenopausal women. Front Horm Res. 1975, 3: 116-130.View ArticlePubMedGoogle Scholar
- Dequeker J, Johnell O: Osteoarthritis protects against femoral neck fracture: the MEDOS study experience. Bone. 1993, 14: S51-56. 10.1016/8756-3282(93)90350-J.View ArticlePubMedGoogle Scholar
- Parfitt AM: Age-related structural changes in trabecular and cortical bone: cellular mechanisms and biomechanical consequences. Calcif Tissue Int. 1984, 36: S123-128. 10.1007/BF02406145.View ArticlePubMedGoogle Scholar
- Ding M, Odgaard A, Linde F, Hvid I: Age-related variations in the microstructure of human tibia cancellous bone. J Orthop Res. 2002, 20: 615-621. 10.1016/S0736-0266(01)00132-2.View ArticlePubMedGoogle Scholar
- Li B, Marshall D, Roe M, Aspden RM: The electron microscope appearance of the subchondral bone plate in the human femoral head in osteoarthritis and osteoporosis. J Anat. 1999, 195: 101-110. 10.1046/j.1469-7580.1999.19510101.x.View ArticlePubMedPubMed CentralGoogle Scholar
- Uhl M, Allmann KH, Tauer U, Laubenberger J, Adler CP, Ihling C: Comparison of MR sequences in quantifying in vitro cartilage degeneration in osteoarthritis of the knee. Br J Radiol. 1998, 71: 291-296.View ArticlePubMedGoogle Scholar
- Gentzsch CM, Jung M, Pueschel K, Delling G, Kaiser E: A scanning electron microscopy based approach toquantify resorption lacunae applied to the trabecular bone of the femural head. J Bone Miner Metab. 2005, 23: 205-211. 10.1007/s00774-004-0585-0.View ArticlePubMedGoogle Scholar
- Gentzsch C, Pueschel K, Deuretzbacher G, Delling G, Kaiser E: First inventory of resorption lacunae on rods and plates of trabecular bone as observed by scanning electron microscopy. Calcif Tissue Int. 2005, 76: 154-162. 10.1007/s00223-004-0212-z.View ArticlePubMedGoogle Scholar
- Dilworth L, Omoruyi FO, Reid W, Asemota HN: Bone and faecal minerals and scanning electron microscopic assessments of femur in rats fed phytic acid extract from sweet potato (Ipomoea batatas). Biometals. 2008, 21: 133-141. 10.1007/s10534-007-9101-z.View ArticlePubMedGoogle Scholar
- Frasca P, Harper RA, Katz JL: Scanning electron microscopy studies of collagen, mineral and ground substance in human cortical bone. Scan Electron Microsc. 1981, 339-346. Pt 3
- Braidotti P, Branca FP, Stagni L: Scanning electron microscopy of human cortical bone failure surfaces. J Biomech. 1997, 30: 155-162. 10.1016/S0021-9290(96)00102-9.View ArticlePubMedGoogle Scholar
- Byers PD, Contepenni CA, Farkas TA: A post mortem study of the hip joint. Ann Rheum Dis. 1970, 29: 15-31. 10.1136/ard.29.1.15.View ArticlePubMedPubMed CentralGoogle Scholar
- Pogrund H, Rutemberg M, Makin M, Robin G, Menczel J, Steiberg R: Osteoarthritis of the hip joint and osteoporosis: a radiological study in a random population sample in Jerusalem. Clin Orthop. 1982, 164: 130-135.PubMedGoogle Scholar
- Li B, Aspden RM: Material properties of bone from the femoral neck and calcar femorale of patients with osteoporosis or osteoarthritis. Osteoporos Int. 1997, 7: 450-456. 10.1007/s001980050032.View ArticlePubMedGoogle Scholar
- Papadakis M, Papadokostakis G, Stergiopoulos K, Kampanis N, Katonis P: Lumbar lordosis in osteoporosis and inosteoarthritis. Eur Spine J. 2009,Google Scholar
- Peyrin F, Salome M, Cloetens P, Laval-Jeantet AM, Ritman E, Ruegsegger P: Micro-CT examinations of trabecular bone samples at different resolutions: 14, 7 and 2 micron level. Technol Health Care. 1998, 6: 391-401.PubMedGoogle Scholar
- Zhang ZM, Jiang LS, Jiang SD, Dai LY: Differentialarticular calcified cartilage and subchondral bone in postmenopausalwomen with osteoarthritis and osteoporosis: two-dimensionalanalysis. Joint Bone Spine.
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