Inflammatory response of intervertebral disc cells is reduced by fibrin sealant scaffold in vitro.

Intervertebral disc (IVD) degeneration is a complex process characterized by elevated concentrations of proinflammatory cytokines and proteolytic enzymes. Because of pro-healing constituents, we hypothesized that fibrin sealant (FS) can reduce inflammation and augment soft tissue healing within the damaged or degenerative IVD. To test this, human and porcine nucleus pulposus (NP) and annulus fibrosus (AF) cells were extracted from tissues and encapsulated into alginate beads (NP cells) and type I collagen sponges (AF cells). Half of the alginate and collagen scaffolds were embedded in FS. To provoke inflammatory behaviours, the constructs were cultured with and without continuous IL-1α (10 ng/ml) for 4, 7 and 14 days. ELISA was used to quantify the cellular synthesis (ng/µg DNA) of clinically relevant cytokines, proteolytic enzymes and growth factors. In NP cell constructs with IL-1α, the syntheses of TNFα, IL-1ß, IL-6, IL-8 was elevated at all culture durations. In the presence of FS, secretion of several pro-inflammatory cytokines were significantly reduced [IL-6 and IL-8 (porcine); and TNFα, IL-1ß, IL-6, IL-8 (human)]. Consistent with these reductions, human NP cultures exposed to FS and FS + IL-1α synthesized significantly reduced amounts of MMP-1 and -3 compared to constructs with IL-1α. For porcine and human AF cells, there were no significant differences in the synthesis of the inflammatory or proteolytic cytokines relative to controls (without IL-1α) at any culture duration. However, the porcine AF cells exposed to FS synthesized elevated amounts of the anti-inflammatory cytokine IL-4. The results suggest that FS may have beneficial effects for patients with degenerated intervertebral discs.

Intradiscal injection of fibrin sealant for the treatment of symptomatic lumbar internal disc disruption: results of a prospective multicenter pilot study with 24-month follow-up.

OBJECTIVE: Assess the safety and efficacy of intradiscal fibrin sealant in adults with chronic discogenic low back pain. DESIGN: Prospective, nonrandomized Food and Drug Administration approved pilot study. SETTING: Three centers in the United States. SUBJECTS: Fifteen adults with chronic, single, or contiguous two-level lumbar discogenic pain confirmed through meticulous provocation discography. INTERVENTIONS: Volume- and pressure-controlled intradiscal delivery of BIOSTAT BIOLOGX(®) Fibrin Sealant with the Biostat(®) Delivery Device into symptomatic lumbar disc(s). OUTCOME MEASURES: Assessments were performed at baseline, 72 hours, and 1, 4, 13, 26, 52, and 104 weeks following intervention. Potential adverse events were evaluated with serial assessment of neurological status, radiographic, and magnetic resonance imaging (MRI). Efficacy measures included serial assessments of low back pain visual analog scale (VAS) measurements and the Roland-Morris Disability Questionnaire (RMDQ). RESULTS: Safety neurological assessments, X-ray, and MRI showed no significant changes. Adverse events were reported in nine subjects. Two instances of low back muscle spasm and one case of discitis were the only events considered related to the procedure or product. EFFICACY: Mean low back pain VAS scores (mm) decreased from 72.4 (95% confidence interval 64.6-80.3) at baseline to 31.7 (17.4-46.1), 35.4 (17.7-53.1), and 33.0 (16.3-49.6); mean RMDQ score improved from 15.2 (12.7-17.7) at baseline to 8.9 (5.3-12.5), 6.2 (3.4-9.1), and 5.6 (2.9-8.4) at 26, 52, and 104 weeks, respectively. CONCLUSION: Intradiscal injection of BIOSTAT BIOLOGX Fibrin Sealant with the Biostat Delivery Device appears safe and may improve pain and function in selected patients with discogenic pain.

Biological and biomechanical effects of fibrin injection into porcine intervertebral discs.

STUDY DESIGN: Surgically denucleated porcine intervertebral discs (IVD) were injected with BIOSTAT BIOLOGX Fibrin Sealant (FS), and the in vivo effects were assessed over time by histological, biochemical, and mechanical criteria. OBJECTIVE: The objectives were to test whether the intradiscal injection of FS stimulates disc healing. SUMMARY OF BACKGROUND DATA: Disc avascularity prevents the deposition of a provisional fibrin scaffold that typically facilitates soft tissue repair. Poor disc wound healing leads to disc damage accumulation and chronic inflammation characterized by overproduction of proinflammatory cytokines and proteolytic enzymes. METHODS: Four lumbar IVDs from each of 31 Yucatan minipigs were randomized to untreated controls; degenerative injury (nucleotomy); and nucleotomy plus FS injection. Animals were killed at 1, 2, 3, 6, and 12 weeks postsurgery. IVDs were harvested to quantify (1) architecture using morphological and histological grading; (2) proteoglycan composition using DMMB assay; (3) cytokine content using ELISA; and (4) mechanical properties using quantitative pressure/volume testing. RESULTS: There was progressive invasion of annular tissue into the nucleus of nucleotomy discs and concomitant reduction in proteoglycan content. By contrast, FS supplementation inhibited nuclear fibrosis and facilitated proteoglycan content recovery over time. FS discs synthesized significantly less TNF-α than degenerate discs (66% vs. 226%, P < 0.05) and had upregulation of IL-4 (310% vs. 166%) and TGF-ß (400% vs. 117%) at 2 to 3 weeks posttreatment. At the third week postsurgery, the denucleated discs were less stiff than controls (pressure modulus 779.9 psi vs. 2754.8 psi; P < 0.05) and failed at lower pressures (250.5 psi vs. 492.5 psi; P < 0.05). The stiffness and leakage pressure of the FS-treated discs recovered to control values after 6 and 12 weeks, respectively. CONCLUSION: FS facilitated structural, compositional, and mechanical repair of the surgically damaged IVD. These FS-derived benefits are likely due to its conductive scaffold properties and metabolically active constituents such as thrombin, factor XIII, and aprotinin acetate.