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Biomedical subjects

Hermann J Bail

Publications and source records attributed to Hermann J Bail.

10 recordsLinked to original sources

CYR61 (CCN1) protein expression during fracture healing in an ovine tibial model and its relation to the mechanical fixation stability.

The formation of new blood vessels is a prerequisite for bone healing. CYR61 (CCN1), an extracellular matrix-associated signaling protein, is a potent stimulator of angiogenesis and mesenchymal stem cell expansion and differentiation. A recent study showed that CYR61 is expressed during fracture healing and suggested that CYR61 plays a significant role in cartilage and bone formation. The hypothesis of the present study was that decreased fixation stability, which leads to a delay in healing, would lead to reduced CYR61 protein expression in fracture callus. The aim of the study was to quantitatively analyze CYR61 protein expression, vascularization, and tissue differentiation in the osteotomy gap and relate to the mechanical fixation stability during the course of healing. A mid-shaft osteotomy of the tibia was performed in two groups of sheep and stabilized with either a rigid or semirigid external fixator, each allowing different amounts of interfragmentary movement. The sheep were sacrificed at 2, 3, 6, and 9 weeks postoperatively. The tibiae were tested biomechanically and histological sections from the callus were analyzed immunohistochemically with regard to CYR61 protein expression and vascularization. Expression of CYR61 protein was upregulated at the early phase of fracture healing (2 weeks), decreasing over the healing time. Decreased fixation stability was associated with a reduced upregulation of the CYR61 protein expression and a reduced vascularization at 2 weeks leading to a slower healing. The maximum cartilage callus fraction in both groups was reached at 3 weeks. However, the semirigid fixator group showed a significantly lower CYR61 immunoreactivity in cartilage than the rigid fixator group at this time point. The fraction of cartilage in the semirigid fixator group was not replaced by bone as quickly as in the rigid fixator group leading to an inferior histological and mechanical callus quality at 6 weeks and therefore to a slower healing. The results supply further evidence that CYR61 may serve as an important regulator of bone healing.

Animals↗

Local cooling restores microcirculatory hemodynamics after closed soft-tissue trauma in rats.

BACKGROUND: Severe closed soft-tissue injury (CSTI) results in progressively developing microvascular dysfunction and local inflammation. Cooling reduces swelling, pain, cellular oxygen demand, and metabolic activity. However, effects of cooling on posttraumatic microcirculation are not yet fully understood. Thus, we assessed effects of local cooling on microcirculation, regional inflammatory response including leukocyte-endothelial cell interaction, and edema formation after CSTI. METHODS: Standardized CSTI was induced by means of controlled impact injury in the left tibial compartment of 14 male Sprague-Dawley rats. Rats were assigned to four groups (n = 7 per group) as follows: group I, no trauma/no cooling; group II, no trauma/20 minutes of cooling; group III, 1.5 hours posttrauma/no cooling; and group IV, 1.5 hours posttrauma/20 minutes of cooling. RESULTS: CSTI resulted in a significant decrease in functional capillary density, a marked increase in microvascular permeability, and granulocyte infiltration (HIS48) as revealed by intravital microscopy and immunohistochemistry of the left extensor digitorum longus muscle. After 20 minutes of local cooling, these microvascular derangements were restored to the level of controls (group I). Edema (extensor digitorum longus muscle wet-to-dry weight ratio) was less pronounced compared with noncooling conditions (group III). Immunoreactivity for HIS48 (neutrophilic granulocytes) in injured rats subjected to local cooling (group IV) was markedly decreased compared with noncooling conditions (group III). CONCLUSION: These results provide in vivo evidence that cooling affords protection of posttraumatic microcirculation through sustained inhibition of microvascular and endothelial dysfunction leading to less granulocyte-dependent inflammation and skeletal muscle edema. Local cooling appears to reduce propagation of acute microvascular injury, preventing leukocyte-dependent tissue destruction and escalation of secondary tissue damage after musculoskeletal soft-tissue trauma.

Animals↗

Instability prolongs the chondral phase during bone healing in sheep.

In this sheep study, we investigated the influence of fixation stability on the temporal and spatial distribution of tissues in the fracture callus. As the initial mechanical conditions have been cited as being especially important for the healing outcome, it was hypothesized that differences in the path of healing would be seen as early as the initial phase of healing. Sixty-four sheep underwent a mid-shaft tibial osteotomy that was treated with either a rigid or a semi-rigid external fixator. Animals were sacrificed at 2, 3, 6 and 9 weeks postoperatively and the fracture calluses were analyzed using radiological, biomechanical and histological techniques. Statistical comparison between the groups was performed using the Mann-Whitney U test for unpaired non-parametric data. In the callus of the tibia treated with semi-rigid fixation, remnants of the fracture haematoma remained present for longer, although new periosteal bone formation during early healing was similar in both groups. The mechanical competence of the healing callus at 6 weeks was inferior compared to tibiae treated with rigid fixation. Semi-rigid fixation resulted in a larger cartilage component of the callus, which persisted longer. Remodeling processes were initiated earlier in the rigid group, while new bone formation continued throughout the entire investigated period in the semi-rigid group. In this study, evidence is provided that less rigid fixation increased the time required for healing. The process of intramembranous ossification appeared during the initial stages of healing to be independent of mechanical stability. However, the delay in healing was related to a prolonged chondral phase.

Animals↗

Initial vascularization and tissue differentiation are influenced by fixation stability.

Fracture healing requires a certain degree of mechanical stability and an adequate blood supply. The hypothesis of the present study was that increased interfragmentary shear leads to a reduced initial vascularization and prolonged healing. The aim of the study was to quantitatively analyze the histological appearance of vascularization and tissue differentiation with regard to fracture stability during the course of healing. A mid-shaft osteotomy of the tibia was performed in two groups of sheep and stabilized with either a rigid or semirigid external fixator, differing in bending stiffness. Interfragmentary movements and ground reaction forces were evaluated in vivo during a 9-week period. The sheep were sacrificed at 2, 3, 6, and 9 weeks postoperatively. The tibiae were tested biomechanically and histological sections from the callus were prepared for analysis of tissue differentiation and vascularization. Larger interfragmentary shear movements in the semirigid fixator group were associated with a reduced initial blood supply. At 6 weeks the semirigid fixator group showed a significantly lower percentage of mineralized bone and a higher amount of fibrous tissue leading to a significantly lower stiffness of the callus than the rigid fixator group. This initial delay in healing was compensated for in the later stages with the production of greater volumes of callus tissue so that both groups showed the same callus stiffness at 9 weeks. However, the rigid fixator group showed signs of the beginning of callus remodeling at the latest time points suggesting a faster bone healing. The results indicate the important role of the initial mechanical stability specifically in the vascularization of an osteosynthesis. Further studies should illustrate the precise role of mechanical conditions on the regulation of angiogenesis during early bone healing.

Animals↗

Three different stages of bone stress reactions of long bones in one patient: case report and review of the literature.

INTRODUCTION: The similar clinical and radiological early course of stress fractures and bone tumors can lead to diagnostic misinterpretation. MATERIALS AND METHODS: A patient is presented who was admitted to our clinic with the diagnosis of similar bone tumors in the distal femur and the proximal tibia. We found an additional localization with high bone turnover in the contralateral proximal tibia; including this lesion, the patient presented with three different types of stress reaction of bone. The final diagnostic decision-making was based on MRI and bone scintigraphy as well as the consideration of localization, age of occurrence, and behavior of malignant bone diseases. Therefore, bone biopsy was not performed. RESULTS: After 8 weeks of partial weight-bearing, the symptoms disappeared, and the patient could return to normal activity.

Adult↗

Endoligamentous revascularization of an anterior cruciate ligament graft.

After replacement of the anterior cruciate ligament with a free tendon graft, the substitute initially is avascular and without a synovial surface. To ensure long-term survival, the graft must become revascularized. Despite numerous studies on the topic, there still is discussion regarding revascularization. The goal of the current study was to investigate the endoligamentous microcapillary revascularization of the free tendon graft after anterior cruciate ligament replacement with time. Thirty-six mature sheep had an anterior cruciate ligament reconstruction with an ipsilateral flexor tendon split graft. Midsubstance tissue samples were immunostained for von Willebrand factor (Factor VIII) at 6, 9, 12, 24, 52, and 104 weeks to detect the endothelial cells of capillaries. Cross sections of vessels were determined in three zones (subsynovial, intermediate, and center of the graft) and were evaluated by means of histomorphometry using a digital imaging analysis system. The observations showed that capillary vessels, which originate from the synovial envelope, invaded the avascular graft tissue from the surface toward the center zone. The highest level of vascular density was found after 6 weeks, reaching the vascular status of the native anterior cruciate ligament after 24 weeks. These findings are in contrast to the findings of previous studies in animals. The current study showed, for the first time, the kinetics of an endoligamentous revascularization of a free tendon graft at the capillary level. In the current model, the process of revascularization terminated earlier than previously described.

Animals↗

Alpha-smooth muscle actin is expressed by fibroblastic cells of the ovine anterior cruciate ligament and its free tendon graft during remodeling.

Contractile fibroblastic cells expressing the alpha-smooth muscle actin isoform, so-called myofibroblasts, have been identified to play a possible role during the healing of the medial collateral ligament by means of restoring the tissues in situ strain via extracellular matrix contraction. Recently, these cells have also been identified to be a normal part of the human anterior cruciate ligament. It has been hypothesized that myofibroblasts play a role in the wrinkling of the extracellular matrix. The goal of the present study was to identify myofibroblasts in the intact ovine anterior cruciate ligament and a free autologous tendon graft during remodeling after anterior cruciate ligament reconstruction. In 36 mature merino sheep the anterior cruciate ligament was replaced with an ipsilateral Achilles tendon split graft. Midsubstance tissue samples were immunostained for alpha-smooth muscle actin at 6, 9, 12, 24, 52, and 104 weeks. Myofibroblasts were identified in the intact ovine anterior cruciate ligament as well as in the Achilles tendon graft prior to implantation. During remodeling the first myofibroblasts were found at six weeks within newly formed fiber bundles. At 24, 52, and 104 weeks myofibroblast distribution and cell density were similar to those of the intact ovine anterior cruciate ligament. These findings indicate that alpha-smooth muscle actin containing fibroblastic cells are a regular part of the intact as well as the remodeled anterior cruciate ligament. There is evidence that myofibroblasts may be involved in maintaining tissue homeostasis in the mature ligament e.g., by means of crimp formation. The presence of these cells during the early remodeling may further indicate that alpha-smooth muscle actin containing fibroblastic cells are involved in the earliest stages of fiber bundle formation. The role and function of this special cell type for the anterior cruciate ligament needs to be further clarified.

Achilles Tendon↗

Tendon healing in a bone tunnel. Part II: Histologic analysis after biodegradable interference fit fixation in a model of anterior cruciate ligament reconstruction in sheep.

PURPOSE: Tendon-to-bone healing of soft-tissue grafts has been described to progress by the development of a fibrous interzone that undergoes a maturation process leading to the development of an indirect type of ligament insertion. Previous studies used extra-articular models or fixation far away from the joint line; thus, no data are available investigating tendon-to-bone healing of a soft-tissue graft fixed anatomically. Therefore, we studied the tendon-to-bone healing of the anatomic soft-tissue graft interference fit fixation in a model of anterior cruciate ligament (ACL) reconstruction in sheep. TYPE OF STUDY: Animal study. METHODS: Thirty-five mature sheep underwent ACL reconstruction with an autologous Achilles tendon split graft. Grafts were directly fixed with biodegradable poly-(D,L-lactide) interference screws. Animals were euthanized after 6, 9, 12, 24, and 52 weeks and histologic evaluations were performed. Undecalcified specimens were evaluated under normal and polarized light. Additionally, animals received a polychrome sequential labeling (tetracycline, xylenol orange, and calcein green) to determine bone growth per time under fluorescent light. RESULTS: Intratunnel histologic findings at 6 weeks showed a tendon-bone junction with only a partial fibrous interzone between the graft tissue and the surrounding bone. A mature intratunnel tendon-bone junction with a zone of fibrocartilage was found at 9 to 12 weeks. At the tunnel entrance site a wide regular ligamentous insertion site was seen in all specimens after 24 weeks. This insertion showed regular patterns such as the direct type of insertion of a normal ligament with a dense basophilic transition zone consisting of mineralized cartilage. CONCLUSIONS: A fibrous interzone between the graft tissue and the bone tunnel was only partially developed, which is in contrast to all previous studies in which nonanatomic fixation was used. Thus, it is reasonable to assume that the tendon-to-bone healing in the present study may progress partially by direct-contact healing without the development of a fibrous interzone. To our knowledge, this is the first report describing the development of a direct type of ligament insertion after ACL replacement with a soft-tissue graft. This is in contrast to previous studies reporting the development of an indirect type of insertion when using nonanatomic fixation far away from the joint line. Thus, histologic data strongly indicate that anatomic interference fit fixation is beneficial for tendon-to-bone incorporation by leading to the development of a direct type of ligament insertion.

Absorbable Implants↗

Ultrasound can predict regenerate stiffness in distraction osteogenesis.

Assessing the regenerate strength in distraction osteogenesis is crucial for clinical treatment. Several methods have been used to achieve this, including plain radiography, digital radiography, quantitative computed tomography, dual energy xray absorptiometry, and ultrasound. The aim of the current study was to investigate the use of ultrasound in monitoring regenerate formation and to correlate this to biomechanical testing results. An osteotomy was done on the tibia of 30 Yucatan micropigs and an Ilizarov-type half-ring external fixator was mounted. After a 5-day latency period, the tibias were distracted for 10 days and then left to consolidate for 10 days. Seven ultrasound examinations were done during Day 15 through Day 25. After sacrifice, the bone mineral density was measured using quantitative computed tomography. Maximum load and torsional stiffness were measured and correlated with ultrasound measurements and bone mineral density. The ultrasound penetration depth and the bone mineral density correlated closely with torsional stiffness. Ultrasound can be a noninvasive predictor of bone regenerate strength in the early phase of distraction osteogenesis, which may reduce the need for radiographs.

Animals↗

Systemic application of growth hormone for enhancement of secondary and intramembranous fracture healing.

Hormones are known to influence bone metabolism and cellular mechanisms of fracture healing. Recent technologies in molecular biology offer recombinant production of hormones, which makes them applicable for pharmacological use. To investigate the effect of systemic growth hormone (GH) application experiments were performed in micropig animal models. Systemic daily subcutaneous injection of species-specific recombinant GH was investigated in Yucatan micropigs to evaluate the effect on secondary fracture healing in a standardized gap model (1 cm) and on intramembranous bone formation in distraction osteogenesis (DO). Quantitative computed tomography (qCT), biomechanical testing, measurement of systemic insulin-like growth factor 1 (IGF-1) levels as well as histomorphometric analyses were performed to investigate differences in regenerate formation. Systemic GH administration significantly increased the torsional stability of the regenerate in comparison to the contralateral side in both experiments. qCT showed accelerated fracture bridging in the GH-treated animals in bone defect healing, while in DO histomorphometry elicited larger callus areas in the case of GH application. Systemic IGF-1 levels were significantly increased in both GH-treated groups. These experiments show that the systemic administration of recombinant GH accelerates fracture healing in standardized animal models. Clinical studies have now been initiated in order to prove the safety and the effectiveness of this therapeutical option.

Animals↗