Mitogenic effects of hydroxyapatite and calcium pyrophosphate dihydrate crystals on cultured mammalian cells

Cheung, Herman S.; Story, Michael T.; McCarty, Daniel J.

doi:10.1002/art.1780270610

Cited by 128 publications

(73 citation statements)

References 22 publications

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“…However, it is important to remember that osteolytic membranes contain a large percentage of both types of cells, and fibroblasts are capable of reacting to particulate debris by secreting a variety of proteases and cytokines which are capable of stimulating bone resorption and osteolysis [32,34]. Other studies have shown that human neonatal fibroblasts have been an excellent model for studying the responsiveness to HA particulates [9,10,12,13,16,20].…”

Section: Discussionmentioning

confidence: 99%

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Effects of hydroxyapatite participate debris on the production of cytokines and proteases in human fibroblasts

Ninomiya

Struve

Stelloh

et al. 2001

Journal Orthopaedic Research

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Cytokines and proteases are secreted by fibroblasts in response to particulate wear debris, and these proteins are felt to play an important role in the development of osteolysis and implant loosening. Although metallic and polyethlyene debris have been studied extensively, little is known about the cellular responses to hydroxyapatite, despite the wide clinical use of these materials. Therefore, the effects of hydroxyapatite (HA) and hydroxyapatitelp-tricalciumphosphate (HARCP) on cellular proliferation, cytokine gene expression and protein secretion, protease synthesis, and gelatinolytic activity were investigated in human fibroblasts.HA and HMTCP particles were synthesized, and their effects were compared to the responses elicited by titanium and cobalt chromium. Sample characterization by scanning electron microscopy and Coulter Counter demonstrated that the materials had a mean particle size of less than 10 pm, and all of the particles were compared using the same concentration ranges.Aliquots of particle suspensions were added to human fibroblasts maintained in tissue culture, and dose-response and timecourse experiments were performed. Effects of the particles on fibroblast proliferation were assessed, and alterations in cytokine levels were determined by specific enzyme linked immunosorbent assays (ELISA). Cytokines that were evaluated included interleukin-1 (IL-1 p), interleukin-6 (IL-6), and tumor necrosis factor-a (TNF-a), all of which have been demonstrated to enhance bone resorption and are associated with osteolysis and implant loosening. Gene expression was determined using Northern blot analysis with cytokine-specific probes, while secretion of the proteases collagenase and stromelysin was determined by Western blot analysis.Functional gelatinolytic assay was assessed using zymogram gels.The particles were evaluated in a concentration range from 0.000021 to 0.021 ~01%. All of the particles produced increases in cellular proliferation up to 0.0021 vol%, with the largest increases being seen at 0.021 vol% with HA/TCP and titanium. At the highest concentration, both cobalt chromium and HA samples decreased cellular proliferation relative to lower doses, possibly representing cytotoxicity.Hydroxyapatite particles yielded a 30-fold increase in interleukin-6 secretion compared to unstimulated controls, which was also greater than three times the levels produced by cobalt chromium, titanium, or HAITCP. HA particles also tripled the secretion of IL-Ip at 0.00021 vol%, and doubled TNF-a secretion at 0.021 ~01%. Addition of conditioned media prepared by incubation of the particles in culture medium in the absence of cells did not alter the secretion of any of the cytokines. Northern blot analysis using IL-6 probes also demonstrated strong increases with HA compared to the other materials, suggesting that the action of the HA particles was at the level of transcription. Secretion of the protease collagenase was increased by all of the samples including HA when compared to unstimulated controls. Stro...

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Section: Discussionmentioning

confidence: 99%

“…Crystals obtained from patients with this syndrome have been shown to induce cellular proliferation and the secretion of prostaglandins and proteases in synovial cells and macrophages [ 11,12,[14][15][16].…”

Section: Introductionmentioning

confidence: 99%

Effects of hydroxyapatite participate debris on the production of cytokines and proteases in human fibroblasts

Ninomiya

Struve

Stelloh

et al. 2001

Journal Orthopaedic Research

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“…A model system of human foreskin fibroblast (HF) cultures was used for these experiments since HF responses to BCP crystals have been shown to be similar to those of synovial fibroblasts (5). HF cultures were established from explants and transferred as described previously (28).…”

Section: Cell Culturementioning

confidence: 99%

“…BCP crystals themselves are at least partly responsible for such proliferation since they stimulate cell replication in vitro (5). Increased cellularity in the synovial lining enhances the capacity for secretion of cytokines, which may promote chondrolysis.…”

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Molecular Mechanism of Basic Calcium Phosphate Crystal-induced Activation of Human Fibroblasts

McCarthy

Augustine

Baldwin

et al. 1998

Journal of Biological Chemistry

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Synovial fluid basic calcium phosphate (BCP) crystals are markers of severe joint degeneration in osteoarthritis. BCP crystals cause mitogenesis of articular cells and stimulate matrix metalloprotease production, thus promoting degradation of articular tissues. Previous work suggested that BCP crystal-induced cell activation required intracellular crystal dissolution, induction of proto-oncogene expression, and activation of signal transduction pathways involving protein kinase C and mitogen-activated protein kinases. Here we further elucidate the mechanisms of BCP crystal-induced cell activation as BCP crystals activate transcription factors nuclear factor B and activator protein 1 in human fibroblasts. We confirm the role of protein kinase C in BCP crystal-induced mitogenesis in human fibroblasts.In contrast, we demonstrate that BCP crystals do not activate signal transduction pathways involving protein tyrosine kinases or phosphatidylinositol 3-kinase. These data further define the mechanism of cell activation by BCP crystals and confirm its selectivity, an observation that may have therapeutic implications. Synovial fluid basic calcium phosphate (BCP)1 (hydroxyapatite, octacalcium phosphate, and tricalcium phosphate) crystals are common in osteoarthritis (OA) and are associated with severe degenerative arthropathies (1). The prevalence of BCP crystals in synovial fluid from patients with knee OA is between 30 and 60%, and their presence correlates strongly with radiographic evidence of cartilage degeneration (2). Larger joint effusions are seen in affected joints when compared with joint fluid from OA knees without BCP crystals (3).Clinical and pathological studies have demonstrated that synovial lining proliferation of varying degrees is associated with BCP crystals in osteoarthritis (4). BCP crystals themselves are at least partly responsible for such proliferation since they stimulate cell replication in vitro (5). Increased cellularity in the synovial lining enhances the capacity for secretion of cytokines, which may promote chondrolysis. Noninflammatory destruction of matrix-rich articular structures including cartilage, ligament, and tendon is also characteristic of BCP crystal deposition (1). BCP crystals promote tissue damage by induction of matrix metalloprotease (MMP) synthesis and secretion. Since there are no available drugs to inhibit deposition or affect reabsorption of these crystals, prevention of the biological consequences of the destructive processes initiated by BCP crystals is an attractive therapeutic strategy.The in vitro effects of BCP crystals emphasize their pathogenic potential. BCP crystals induce mitogenesis in cultured fibroblasts (5, 6). However, mechanisms by which BCP crystals induce mitogenesis have been incompletely studied. Endocytosis and intracellular dissolution of the crystals, producing an elevated cytoplasmic calcium concentration, are important (7). BCP crystals activate a protein kinase signal transduction pathway involving p42 and p44 mitogen-activated protein (MA...

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Differential effects of different forms of hydroxyapatite and hydroxyapatite/tricalcium phosphate particulates on human monocyte/macrophagesin vitro

Harada

Wang

Doppalapudi

et al. 1996

J. Biomed. Mater. Res.

100

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A possible complication associated with the use of hydroxyapatite (HA) or HA/tricalcium phosphate (HA/TCP) coating on the surfaces of prosthetic devices used for dental and orthopedic implants is their potential to fragment and thus exist as wear debris. In contrast to the so-called osteoconductive properties of HA or HA/TCP coatings, in particulate form these materials may lead to an adverse pattern of cellular and tissue responses at the bone-implant interface. We have established an in vitro cell culture system to characterize the biologic and biochemical effects of various particulate materials. The present study demonstrates that the HA/TCP particles derived from different sintering temperatures exhibit differential effects on cultured human monocyte/macrophages (M/M). The HA/TCP particles dried at 110 degrees C were the most biologically active, stimulating significant release of interleukin 1 beta (IL-1 beta), IL-6, tumor necrosis factor alpha, and prostaglandin E2 (PGE2), products implicated as important mediators of inflammation in diverse pathologic conditions. Other particles, sintered at either 900 or 1200 degrees C, did not stimulate production of cytokines or PGE2. HA/TCP particles from plasma-spray coatings also failed to release proinflammatory products. These results suggest that the biochemical and crystalline structural properties of particles markedly affects their capacity to modulate M/M function. This in vitro culture system should be useful in characterizing the specific physical and chemical properties of HA or HA/TCP particulates that are responsible for stimulating proinflammatory cell responses.

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Mitogenic effects of hydroxyapatite and calcium pyrophosphate dihydrate crystals on cultured mammalian cells

Cited by 128 publications

References 22 publications

Effects of hydroxyapatite participate debris on the production of cytokines and proteases in human fibroblasts

Effects of hydroxyapatite participate debris on the production of cytokines and proteases in human fibroblasts

Molecular Mechanism of Basic Calcium Phosphate Crystal-induced Activation of Human Fibroblasts

Differential effects of different forms of hydroxyapatite and hydroxyapatite/tricalcium phosphate particulates on human monocyte/macrophagesin vitro

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