Modification of Mechanical Properties, Polymerization Temperature, and Handling Time of Polymethylmethacrylate Cement for Enhancing Applicability in Vertebroplasty.

Polymethylmethacrylate (PMMA) bone cement is a popular bone void filler for vertebroplasty. However, the use of PMMA has some drawbacks, including the material's excessive stiffness, exothermic polymerization, and short handling time. This study aimed to create an ideal modified bone cement to solve the above-mentioned problems. Modified bone cements were prepared by combining PMMA with three different volume fractions of castor oil (5%, 10%, and 15%). The peak polymerization temperatures, times to achieve the peak polymerization temperature, porosities, densities, modulus and maximum compression strengths of standard (without castor oil), and modified cements were investigated following storage at ambient temperature (22°C) or under precooling conditions (3°C). Six specimens were tested in each group of the aforementioned parameters. Increasing castor oil content and precooling treatment effectively decreased the peak polymerization temperatures and increased the duration to achieve the peak polymerization temperature (P < 0.05). Furthermore, the mechanical properties of the material, including density, modulus, and maximum compression strength, decreased with increasing castor oil content. However, preparation temperature (room temperature versus precooling) had no significant effect (P > 0.05) on these mechanical properties. In conclusion, the addition of castor oil to PMMA followed by precooling created an ideal modified bone cement with a low modulus, low polymerization temperature, and long handling time, enhancing its applicability and safety for vertebroplasty.

Hyperbaric oxygen treatment suppresses MAPK signaling and mitochondrial apoptotic pathway in degenerated human intervertebral disc cells.

Nucleus pulposus cells (NPCs) from degenerating discs produce catabolic and inflammatory factors, including interleukin (IL)-1 and nitric oxide (NO). Enhanced production of NO has been implicated in the apoptosis of degenerating disc cells. This study evaluates the effects of hyperbaric oxygen (HBO) on degenerated human NPCs. All hyperoxic cells were exposed to 100% O(2) at 2.5 atmospheres absolute (ATA). Phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (MAPK) in NPCs was detected using the phosphor-kinase array kit. RNA was isolated for real-time polymerase chain reaction (PCR) analysis of aggrecan and type II collagen gene expression. The levels of IL- 1ß and NO were quantified by enzyme-linked immunosorbent assay (ELISA). To identify the HBO-induced anti-apoptotic pathways, expression of Bcl-2 and Bax proteins as well as activation of cysteine-containing aspartate-specific proteases (caspases) 3, 8, and 9 was evaluated using Western blotting after HBO treatment. Our data showed that HBO treatment decreased the expression of IL-1ß, suppressed phosphorylation of ERK1/2, JNK, and p38 MAPK, decreased synthesis of NO, and increased the gene expression of aggrecan and type II collagen in NPCs as compared with the atmospheric treatment. HBO up-regulated the ratio of Bcl-2 to Bax expression and reduced the activity of caspases 9 and 3 but not of caspase 8, indicating a selective effect over the mitochondrial apoptosis pathway in degenerated NPCs. These results support our hypothesis that HBO treatment suppresses MAPK signaling and mitochondrial apoptotic pathway in degenerated human intervertebral disc cells.

Beneficial effects of hyperbaric oxygen on human degenerated intervertebral disk cells via suppression of IL-1ß and p38 MAPK signal.

Nucleus pulposus cells (NPCs) from degenerating disks produce catabolic and inflammatory factors, including interleukin (IL)-1, nitric oxide (NO), prostaglandin E2 (PGE-2), and matrix metalloproteinaes (MMPs). An imbalance between MMPs and tissue inhibitors of matrix metalloproteinases (TIMPs) has been proposed to exist in the degenerating disk. This study evaluates the effects of hyperbaric oxygen (HBO) on the human degenerated NPCs. NPCs were maintained in alginate bead culture. All hyperoxic cells were exposed to 100% O(2) at 2.5 atmospheres absolute (ATA) in a hyperbaric chamber. p38 MAPK phosphorylation of the NPCs was detected using the phosphor-kinase array kit. RNA was isolated for real-time quantitative polymerase chain reaction (Q-PCR) analysis of aggrecan and type II collagen gene expression. The amounts of IL-1ß, NO, PGE-2, MMP-3, and TIMP-1 in the conditioned media were quantified by enzyme-linked immunosorbent assay (ELISA). Our data showed that HBO treatment decreased expression of IL-1ß, increased the gene expression of aggrecan and type II collagen, suppressed the phosphorylation of p38 MAPK, decreased NO, PGE-2, and MMP-3, and increased TIMP-1 expression in NPCs as compared with the atmospheric treatment. These results support the hypothesis that IL-1ß and the p38 MAPK signal may be responsible for many of the inflammatory and catabolic changes seen in the human disk degeneration, and support our proposal that HBO treatment-induced increase of the anabolic factor (TIMP-1)/catabolic factor (MMP-3) ratio may provide a therapeutic approach to slow the course of intervertebral disk degeneration.

Effect of hyperbaric oxygen on mesenchymal stem cells for lumbar fusion in vivo.

BACKGROUND: Hyperbaric oxygen (HBO) therapy has been proved in improving bone healing, but its effects on mesenchymal stem cells (MSCs) in vivo is not clear. The aims of this study are to clarify whether the HBO therapy has the same enhancing effect on MSCs with regard to bone formation and maturation and to ascertain whether the transplanted MSCs survive in the grafted area and contribute to new bone formation. METHODS: Twenty-three adult rabbits underwent posterolateral fusion at L4-L5 level. The animals were divided into three groups according to the material implanted and subsequent treatment: (1) Alginate carrier (n = 6); (2) Alginate-MSCs composite (n = 11); and (3) Alginate-MSCs composite with HBO therapy (n = 6). After 12 weeks, spine fusion was examined using radiographic examination, manual testing, and histological examination. Using a PKH fluorescence labeling system, whether the transplanted MSCs survived and contributed to new bone formation in the grafted area after HBO therapy was also examined. RESULTS: The bilateral fusion areas in each animal were evaluated independently. By radiographic examination and manual palpation, union for the Alginate, Alginate-MSCs, and Alginate-MSCs-HBO groups was 0 of 12, 10 of 22, and 6 of 12 respectively. The difference between the Alginate-MSCs and Alginate-MSCs-HBO groups was not significant (P = 0.7997). The fluorescence microscopy histological analysis indicated that the transplanted PKH67-labeled MSCs survived and partly contributed to new bone formation in the grafted area. CONCLUSIONS: This study demonstrated that the preconditioned MSCs could survive and yield bone formation in the grafted area. HBO therapy did not enhance the osteogenic ability of MSCs and improve the success of spine fusion in the rabbit model. Although there was no significant effect of HBO therapy on MSCs for spine fusion, the study encourages us to research a more basic approach for determining the optimal oxygen tension and pressure that are required to maintain and enhance the osteogenic ability of preconditioned MSCs. Further controlled in vivo and in vitro studies are required for achieving a better understanding of the effect of HBO treatment on MSCs.

A comparison of posterolateral lumbar fusion comparing autograft, autogenous laminectomy bone with bone marrow aspirate, and calcium sulphate with bone marrow aspirate: a prospective randomized study.

STUDY DESIGN: A prospective clinical study. OBJECTIVE: To evaluate whether the fusion rate of autogenous laminectomy bone chips and calcium sulfate pellets could be augmented by bone marrow aspirate (BMA) in one-level lumbar posterolateral fusion. SUMMARY OF BACKGROUND DATA: An in vivo animal study has indicated that BMA augments spinal arthrodesis. METHODS: Forty-three patients undergoing surgery for instrumented one-level fusion with decompression were divided into 2 groups. Autologous iliac crest bone graft (ICBG) was placed in 1 posterolateral gutter (control), while on the other side (test), an equal quantity of laminectomy bone chips mixed with BMA while harvesting the iliac bone graft (group 1) or an equal quantity of calcium sulfate pellets soaked in BMA (group 2) was placed. Radiographic assessment was performed every 3 months (3-12 months) and then annually. The statuses of fusion on either side of the vertebra were compared. RESULTS: For the 21 patients in group 1, 18 (85.7%) exhibited bone fusion on the test side, and 19 (90.5%) presented evidence of fusion on the control side. Thus, the test side with laminectomy bone chips and BMA achieved a fusion rate similar to that on the control side (P > 0.05). For the 22 patients in group 2, 20 (90.9%) exhibited bone fusion on the control side whereas only 10 (45.5%) demonstrated complete fusion on the test side (P < 0.05), where calcium sulfate and BMA was applied. CONCLUSION: ICBG performs as expected with high fusion rates and laminectomy bone with BMA performs equally as well. Osteoset is significantly inferior to ICBG despite the addition of BMA, which is osteoinductive and has improved fusion rates and osteogenesis in other models.

The fusion rate of calcium sulfate with local autograft bone compared with autologous iliac bone graft for instrumented short-segment spinal fusion.

STUDY DESIGN: A prospective study. OBJECTIVES: To compare the efficacy of calcium sulfate pellets plus laminectomy bone chips with a fresh autologous iliac bone graft for short-segment lumbar fusion. SUMMARY OF BACKGROUND DATA: Bone graft substitute material can be used to expand an existing quantity of available laminectomy bone chips. METHODS: Seventy-four patients underwent surgery for instrumented one- or two-segment fusion with decompression. Autologous iliac crest bone graft was placed in one posterolateral gutter, while on the other side, an equal quantity of autogenous laminectomy bone supplemented with calcium sulfate was placed. Radiographic assessment included radiographs alone; this was performed every 3 months (3 months to 12 months), then annually. The status of fusion and the relative size of the fusion bone mass on either side of the vertebra were compared. RESULTS: Using iliac crest bone graft (control side) versus autograft laminectomy bone with calcium sulfate (test side), there was no significant difference between the fusion rate and sizes of the fusion bone mass (P > 0.05). Follow-up periods ranged from 30 months to 34 months, averaging 32.5 months. For the 39 patients who received single-segment fusion, 34 patients (87.2%) exhibited bone fusion on the test side, and 35 patients (89.7%) had evidence of fusion on the control side. For the 35 patients who received two-segment fusion, 29 patients (82.9%) exhibited bone fusion on the test side and 30 patients (85.7%) demonstrated complete fusion on the control side. CONCLUSIONS: The fusion rate and fusion size between the two groups are similar. Calcium sulfate pellets may play a role as a bone graft extender in short-segment spinal fusion.