Four-year follow-up of poly-L-lactic Acid cages for lumbar interbody fusion in goats.

BACKGROUND: New applications of bioabsorbable polymer implants demand for histologic evaluation because a host tissue response is elicited and late complications after polymer implantation have been reported. Furthermore, in load-bearing regions an accelerated polymer degradation and foreign body reaction may be observed. METHODS: Lumbar interbody fusion procedures were performed using poly-L-lactic acid (PLLA) and titanium cages in 43 goats. At 3, 6, 12, 24, 36, and 48 months after surgery, sequential histologic analysis of instrumented motion segments, lymph nodes, and nervous structures was performed. Blood samples were retrieved for laboratory analysis. RESULTS: No adverse local or distant histologic or systemic effects were observed during the absorption of the poly-L-lactic acid cages. Interbody fusion was maintained, and only a very mild inflammatory response was observed. In half the specimens complete absorption was observed, and in the remaining specimens an estimated 1-10% of the original PLLA was present at the 3-year follow-up. At the 4-year follow-up, five out of seven PLLA specimens showed no PLLA particles under polarized light microscopy. In the remaining two specimens an estimated 1% of the original PLLA could be observed. CONCLUSIONS: Poly-L-lactic acid cages are feasible for lumbar interbody fusion, and the biocompatibility under high load bearing conditions is excellent during the complete absorption of the PLLA interbody fusion cages.

Localisation of osteogenic and osteoclastic cells in porous beta-tricalcium phosphate particles used for human maxillary sinus floor elevation.

We and others have shown earlier that porous beta-tricalcium phosphate (TCP) (Cerasorb) can be used in patients to augment the maxillary sinus floor prior to placement of oral dental implants. To better understand the transformation of TCP particles into bone tissue, we analyse here the appearance of cells with osteogenic or osteoclastic potential in relation to these particles. In biopsies taken at 6 months after sinus floor augmentation we observed bone growth into the TCP particles but also replacement by soft connective tissue. To identify possible osteoprogenitor cells in this tissue, histological sections were immunostained with an antibody to Runx2/Cbfa1, an essential and early transcription factor for osteoblast differentiation. The osteogenic potential of cells was further confirmed by immunostaining for bone sialoprotein (BSP) and osteopontin (OPN). Other sections were stained for Tartrate Resistant Acid Phosphatase (TRAP) activity to identify cells with osteoclastic capacity. Runx2/Cbfa1 positive connective tissue cells were found in abundance throughout and around the TCP particles, even at a distance of several millimetres from the maxillary bone surface. About 95% of the cells found within TCP particles stained positive for Runx2/Cbfa1. Fewer cells stained positive for BSP and OPN, suggesting more mature osteoblastic properties. Mono- and binucleate TRAP-positive cells, but no multinucleate TRAP-positive osteoclasts, were found in the soft tissue infiltrating the TCP and at the surface of the TCP particles. Both the Runx2/Cbfa1 positive and the TRAP-positive cells decreased apically with increasing vertical distance from the maxillary bone surface. This data suggests that the TCP particles attract osteoprogenitor cells that migrate into the interconnecting micropores of the bone substitute material by 6 months. The lack of large multinucleate TRAP positive cells suggests that resorption of the TCP material by osteoclasts plays only a minor role in its replacement by bone. Chemical dissolution, possibly favoured by a high cell metabolism in the particles, seems the predominant cause of TCP degradation. The abundance of Runx2/Cbfa1 positive cells would indicate that with a greater time of healing there will be further bone deposition into these particles.

Strain-derived canalicular fluid flow regulates osteoclast activity in a remodelling osteon--a proposal.

The concept of bone remodelling by basic multicellular units is well established, but how the resorbing osteoclasts find their way through the pre-existing bone matrix remains unexplained. The alignment of secondary osteons along the dominant loading direction suggests that remodelling is guided by mechanical strain. This means that adaptation (Wolff's Law) takes place throughout life at each remodelling cycle. We propose that alignment during remodelling occurs as a result of different canalicular flow patterns around cutting cone and reversal zone during loading. Low canalicular flow around the tip of the cutting cone is proposed to reduce NO production by local osteocytes thereby causing their apoptosis. In turn, osteocyte apoptosis could be the mechanism that attracts osteoclasts, leading to further excavation of bone in the direction of loading. At the transition between cutting cone and reversal zone, however, enhanced canalicular flow will stimulate osteocytes to increase NO production, which induces osteoclast retraction and detachment from the bone surface. Together, this leads to a treadmill of attaching and detaching osteoclasts in the tip and the periphery of the cutting cone, respectively, and the digging of a tunnel in the direction of loading.

Interactive effects of PTH and mechanical stress on nitric oxide and PGE2 production by primary mouse osteoblastic cells.

Parathyroid hormone (PTH) and mechanical stress both stimulate bone formation but have opposite effects on bone resorption. PTH increased loading-induced bone formation in a rat model, suggesting that there is an interaction of these stimuli, possibly at the cellular level. To investigate whether PTH can modulate mechanotransduction by bone cells, we examined the effect of 10-9 M human PTH-(1-34) on fluid flow-induced prostaglandin E2 (PGE2) and nitric oxide (NO) production by primary mouse osteoblastic cells in vitro. Mechanical stress applied by means of a pulsating fluid flow (PFF; 0.6 +/- 0.3 Pa at 5 Hz) stimulated both NO and PGE2 production twofold. In the absence of stress, PTH also caused a twofold increase in PGE2 production, but NO release was not affected and remained low. Simultaneous application of PFF and PTH nullified the stimulating effect of PFF on NO production, whereas PGE2 production was again stimulated only twofold. Treatment with PTH alone reduced NO synthase (NOS) enzyme activity to undetectable levels. We speculate that PTH prevents stress-induced NO production via the inhibition of NOS, which will also inhibit the NO-mediated upregulation of PGE2 by stress, leaving only the NO-independent PGE2 upregulation by PTH. These results suggest that mechanical loading and PTH interact at the level of mechanotransduction.

The use of poly-L-lactic acid in lumbar interbody cages: design and biomechanical evaluation in vitro.

Cage design and cage material may play a crucial role in the incidence of postoperative complications reported with current non-absorbable interbody cage devices. Bioabsorbable poly-L-lactic acid cage devices may have potential benefits. The purpose of this study was to determine the required strength of poly-L-lactic acid cages for use in experimental goat studies and to evaluate the mechanical properties of different cage designs in situ. The yield and ultimate strength of native goat motion segments (L1-L6) were determined; the yield strength was used as a design parameter for the cages. The mechanical behaviour of two types of poly-L-lactic acid cages, the influence of endplate perforation, differences between toothed and smooth cages, and the influence of cage filling were biomechanically tested and compared to native motion segments. Only axial compression until failure of the motion segments was performed. Dual energy X-ray absorptiometry was used to determine bone mineral content. The yield and ultimate strength of the native motion segments were 3.5 and 7.0 kN, respectively. Based on these data, flexible and stiff poly-L-lactic acid cages were designed with strengths of 3.5 and 7 kN, respectively. Poly-L-lactic acid cages, whether with or without bone graft and perforating the endplates, did not reduce the compressive strength of motion segments as compared to native segments. However, toothed titanium cages, with the same geometry, negatively influenced the segments' compressive strength, which effect was reduced using smooth titanium cages.

Osteocyte and bone structure.

The osteocyte is the most abundant cell type of bone. There are approximately 10 times as many osteocytes as osteoblasts in adult human bone, and the number of osteoclasts is only a fraction of the number of osteoblasts. Our current knowledge of the role of osteocytes in bone metabolism is far behind our insight into the properties and functions of the osteoblasts and osteoclasts. However, the striking structural design of bone predicts an important role for osteocytes in determining bone structure. Over the past several years, the role of osteocytes as the professional mechanosensory cells of bone, and the lacunocanalicular porosity as the structure that mediates mechanosensing have become clear. Strain-derived flow of interstitial fluid through this porosity seems to mechanically activate the osteocytes, as well as ensure transport of cell signaling molecules, nutrients, and waste products. This concept explains local bone gain and loss--as well as remodeling in response to fatigue damage--as processes supervised by mechanosensitive osteocytes. Alignment during remodeling seems to occur as a result of the osteocyte's sensing different canalicular flow patterns around the cutting cone and reversal zone during loading, therefore determining the bone's structure.

Fate of monocortical bone blocks grafted in the human maxilla: a histological and histomorphometric study.

Local bone defects in the anterior maxilla are commonly grafted with monocortical blocks of autologous bone in order to restore the defect site prior to the placement of dental implants. Increasing evidence suggests that osteocytes are involved in the control of bone remodelling and thus may be important for optimalisation of bone structure around implants, and thus for implant osseointegration. However, it is not well known whether osteocytes will survive when bone blocks are grafted into defects. We grafted 19 patients with monocortical bone blocks derived from the symphysis, to the defect site in the maxillary alveolar process. The bone grafts were left to heal for times varying from 2.5 to 7 months. During implant installation, bone biopsies were removed using a trephine burr, and processed for hard tissue histology. Bone histology and histomorphometry were then carried out in order to gain insight into the density, viability and remodelling of the graft. Clinically, all the bone grafts were successful, with no implant failures, and little resorption was seen. Histologically, bone volume expressed as percentage of tissue volume at the implant site varied from 27% to 57% with an overall average of 41%. Bone fields with empty osteocyte lacunae were observed and measured. The amount of this so-called nonvital bone (NVB) varied between 1% and 34% of the total tissue volume. The amount of NVB decreased significantly with the time of healing. The data suggest that the majority of the osteocytes of the monocortical bone do not survive grafting. The results indicate that the NVB is progressively remodelled into new vital bone 7 months after grafting.

Bioabsorbable poly-L-lactic acid cages for lumbar interbody fusion: three-year follow-up radiographic, histologic, and histomorphometric analysis in goats.

STUDY DESIGN: Long-term evaluation was performed for bioabsorbable poly-L-lactic acid cages in a goat interbody fusion model. OBJECTIVE: To assess the radiographic, histologic, and histomorphometric characteristics of poly-L-lactic acid cages during 3 years of follow-up evaluation. SUMMARY OF BACKGROUND DATA: Failed cage fusions may be related to cage design and material in addition to the surgical technique used. To overcome material-related complications and to explore the potential benefits of bioabsorbable cages, poly-L-lactic acid cages have been designed. METHODS: For this study, 36 Dutch milk goats underwent a lumbar interbody fusion procedure (L3-L4). Two types of custom-made cage devices were impacted with bone graft and implanted: poly-L-lactic acid cages (n = 30) and titanium cages (n = 6). Sequential harvesting of surgically managed motion segments (intervals: 3, 6, 12, 24, and 36 months) was performed for analysis. RESULTS: In poly-L-lactic acid specimens, permanent interbody fusion could be achieved within 6 months after surgery with maintenance of cage height. Titanium specimens showed no interbody fusion within this period. Radiographic follow-up evaluation (6-36 months) showed interbody fusion in 86% (19/22) of poly-L-lactic acid specimens, as compared with 33% (2/6) of titanium specimens. After 36 months of implantation, in one half of the specimens, poly-L-lactic acid cages were completely absorbed. Bone histomorphometry showed complete bone remodeling after 2 years of follow-up evaluation. During the study period, no local or distant adverse histologic effects were observed. CONCLUSIONS: The current study showed that poly-L-lactic acid cage devices are feasible for lumbar interbody fusion. New poly-L-lactic acid cages designed for clinical practice might be a viable alternative to current nonabsorbable cage devices.

A case for strain-induced fluid flow as a regulator of BMU-coupling and osteonal alignment.

Throughout life, human bone is renewed continuously in a tightly controlled sequence of resorption and formation. This process of bone remodeling is remarkable because it involves cells from different lineages, collaborating in so-called basic multicellular units (BMUs) within small spatial and temporal boundaries. Moreover, the newly formed (secondary) osteons are aligned to the dominant load direction and have a density related to its magnitude, thus creating a globally optimized mechanical structure. Although the existence of BMUs is amply described, the cellular mechanisms driving bone remodeling-particularly the alignment process-are poorly understood. In this study we present a theory that explains bone remodelling as a self-organizing process of mechanical adaptation. Osteocytes thereby act as sensors of strain-induced fluid flow. Physiological loading produces stasis of extracellular fluid in front of the cutting cone of a tunneling osteon, which will lead to osteocytic disuse and (continued) attraction of osteoclasts. However, around the resting zone and the closing cone, enhanced extracellular fluid flow occurs, which will activate osteocytes to recruit osteoblasts. Thus, cellular activity at a bone remodeling site is well related to local fluid flow patterns, which may explain the coordinated progression of a BMU.

High concentrations of bioactive glass material (BioGran) vs. autogenous bone for sinus floor elevation.

In this study, high concentrations of bioactive glass (BG) particles were compared with autogenous bone in their capacity to augment maxillary bone when grafted in the human sinus floor using a split mouth design. Three female patients with severe maxillary atrophy underwent bilateral sinus floor elevation and bone grafting using 80-100% BG particles (300-355 microm in size) mixed with 20% to 0% iliac crest bone particles at one (experimental) side, and 100% iliac crest derived bone particles at the other (control) side. A total of 22 bone biopsies was taken at the time of fixture installation; that is, at 4, 6 and 15 months after grafting, and processed for histology and histomorphometry. At the control (autogenous bone) sides, trabecular bone amounted to 39% of the biopsy volume in the graft (site) at 4 months, almost 41% at 6 months, and 42% at 15 months. This bone contained viable osteocytes and was mostly of mature, lamellar type. At the experimental (BG particles) sides, the graft consisted of 27% of mostly woven (and some lamellar) bone at 4 months, 36% (woven and lamellar) bone at 6 months, and 39% (mainly lamellar) bone at 15 months. The grafted BG particles started to excavate at 4 months and their centers gradually filled with bone tissue. As a consequence, the volume of BG particles in the biopsy decreased from 29% at 4 months to 15% at 6 months and 8% at 15 months. The BG particles appeared to resorb within 1-2 years by dissolution rather than by osteoclastic activity. Parameters for bone turnover (% osteoid surface, % resorption surface) indicated that bone remodeling was very active at both experimental and control sides, during more than 6 months. These results suggest that mixtures of mainly (80-90%) BG particles and some (10-20%) autogenous bone are effective for bone regeneration in the augmented sinus offer 6 months healing time, while about 12 months healing time is needed for 100% BG particles.

The effect of cage stiffness on the rate of lumbar interbody fusion: an in vivo model using poly(l-lactic Acid) and titanium cages.

STUDY DESIGN: A goat interbody fusion model using poly-(L-lactic acid) and titanium cages was designed to evaluate the effect of cage stiffness on lumbar interbody fusion. OBJECTIVE: To investigate the effect of cage stiffness on the rate of interbody fusion. SUMMARY OF BACKGROUND DATA: Various types of cages considerably exceed the stiffness of vertebral bone, which ultimately may lead to postoperative complications. To avoid these complications, poly-(L-lactic acid) cages with limited stiffness have been designed. The mechanical integrity of the cages remains intact for at least 6 months. METHODS: Interbody fusions were performed at L3-L4 of 15 Dutch milk goats, and one of three cages was randomly implanted: 1) a titanium cage (n = 3), 2) a stiff poly-(L-lactic acid) cage (n = 6), or 3) a flexible poly-(L-lactic acid) cage (n = 6). Interbody fusion was assessed radiographically by three independent observers 3 and 6 months after surgery. RESULTS: At 3 months, all the poly-(L-lactic acid) specimens showed ingrowth of new bone, but with radiolucency in the fusion mass. At 6 months, solid arthrodesis was observed in four of six poly-(L-lactic acid) specimens, advanced ingrowth in one specimen, and infection in one specimen. Titanium cages showed ingrowth of bone, but with radiolucency in the fusion mass. Interbody fusion using poly-(L-lactic acid) cages showed a significantly higher rate statistically (P = 0.016) and more complete fusion than titanium cages of the same design. CONCLUSIONS: The reduced stiffness of poly-(L-lactic acid) cages showed enhanced interbody fusion, as compared with titanium cages after 6 months. Bioabsorbable poly-(L-lactic acid) cages thus may be a viable alternative to current interbody cage devices, thereby avoiding the concomitant problems related to their excessive stiffness. However, the bioabsorbability of the poly-(L-lactic acid) cages awaits investigation in a long-term study currently underway.