The temporalis myo-osseous flap: an experimental study

The present paper investigates the anatomy and vascularization of the temporalis myo-osseous flap. This is a calvarial bone flap that employs temporalis muscle and its distal pericranial extension as a pedicle. In six human cadavers the flap was raised as an island on the anterior deep temporal artery after transecting the zygomatic arch and coronoid process. Maximal mobilization was thus obtained, allowing rotation of the flap into the mouth for intraoral reconstruction. The arc of rotation and potential surgical applications were noted. A comparative study of the temporalis myo-osseous flap and free calvarial bone graft was then conducted in a rabbit model. Vascularization of the calvarial bone flap was confirmed by technetium scintigraphy performed on the first postoperative day. The uptake of fluorochrome labels immediately after transfer verified the adequacy of the periosteal circulation in maintaining viability and new osteoid formation throughout the full thickness of calvarial bone. The transplantation of free calvarial bone grafts was followed by necrosis of most cellular elements. This was demonstrated by an absence of fluorochrome uptake up to 19 days postoperatively and a predominance of empty lacunae and nonviable marrow.     

Use of the temporalis muscule flap in blanking out orbits.

We report the use of the temporalis muscle as a transposition flap to obliterate the orbit in 5 patients. In 4 of the cases we split the muscle coronally and passed the anterior part through a window in the lateral orbital wall. In two of these patients, skin grafts were put on both sides of the temporalis muscle-fascia flap, to restore nasal lining and to cover the facial surface simultaneously. In the remaining patients, the muscle was split sagittally to provide a large surface for coverage. The temporalis muscle flap is a versatile one for filling orbits after exenteration.     

Reconstruction of the burned ear using a temporalis fascial flap

A case report is presented in which a temporalis fascial flap is used to reconstruct a burned ear complicated by full-thickness skin loss with exposed cartilage. The method provides a simple, reliable means for covering exposed auricular cartilage with readily obtainable vascularized tissue. Donor-site morbidity is minimal because the scar lies within the hairline.     

The blood supply of the reverse temporalis muscle flap: anatomic study and clinical implications

Although the reverse temporalis muscle flap has been used clinically, the exact vascular connection between the superficial and deep temporal vessels has not been clearly defined. The purpose of this study was to investigate the vascular territory of the reverse temporalis muscle supplied by the superficial temporal vessels. Six cadaver heads were studied using a colored lead oxide injection through the superficial temporal artery. The specimens were examined macroscopically and radiographically. The reverse temporalis muscle flap was then applied to a clinical case presenting with traumatic anterior skull base defect communicating with the nasal cavity. The cadaver specimens demonstrated that the superficial temporal artery formed an average 1.3 +/- 0.2 cm in width of dense vascular zone, which was located within 1.8 cm below the superior temporal line. The dense vascular network further perfused the anterior and posterior deep temporal arteries and the muscular branch of the middle temporal artery to supply the temporalis muscle. The mean perfused area of the temporalis muscle was 83 percent, ranging from 79 to 89 percent, in five cadaver heads. One cadaver revealed only 55 percent of perfused area in the absence of the muscular branch of the middle temporal artery. The consistent area without perfusion was located in the distal third of the posterior portion of the reverse temporalis muscle. In clinical cases, the reverse temporalis muscle flap was used successfully to obliterate the anterior skull base defect without evidence of muscle flap necrosis. The exact blood supply to the distal third of the posterior portion of the reverse temporalis muscle flap needs to be investigated further in vivo. Particular attention was paid to the inclusion of the muscular branch of the middle temporal artery in this flap to augment the blood supply to the temporalis muscle.     

Cyclooxygenase-2 inhibition does not impair block bone grafts healing in rabbit model

Success of alveolar reconstructions using onlay autogenous block bone grafts depends on their adequate integration to the recipient bed influenced by a number of local molecules. Considering the fundamental role of cyclooxygenase (COX-2) in bone repair, the aim of this study was to analyze the effect of its inhibition in the integration of endochondral (EC) iliac crest, and intramembranous (IM) calvaria bone grafts. Thirty-two rabbits were divided into 4 groups: Calvaria Control (CC) and Iliac Control—treated with oral 0.9 % saline solution, and Calvarial-NSAID (C-NSAID) and Iliac-NSAID (I-NSAID) groups—treated with oral 6 mg/Kg non-steroidal anti-inflammatory drug etoricoxib. After 7, 14, 30 and 60 days the animals were euthanized and the specimens removed for histological, histomorphometric and immunohistochemistry analysis. At day 60, a tight integration of IM blocks could be seen with the presence of remodeling bone, whereas integration of EC grafts was mainly observed at the edges of the grafts. A significant higher percentage of bone matrix in the interface region of the CC grafts in comparison to C-NSAID only at day 14, whereas no differences were detected comparing the EC grafts. No differences were observed in Runx-2 and vascular endothelial growth factor (VEGF) immunolabeling when comparing CC and C-NSAID groups, while a significant weaker Runx-2 and VEGF labeling was detected in I-NSAID group at day 60. Although some influence was detected in osteogenesis, it is concluded that drug induced inhibition of COX-2 does not impair onlay bone grafts’ healing of both embryologic origins in rabbits.     

Repair of the pharyngocutaneous fistula with a fasciocutaneous island flap pedicled on the superficial temporalis artery

The case of an 84-year-old man with a pharyngocutaneous fistula after radiotherapeutic treatment and total laryngectomy for a squamous cell carcinoma was reported. Treatment with local flaps failed and normal flaps were not likely to succeed because of general and locally poor conditions considering that intensive radiotherapy had been administered. We therefore decided to use a fasciocutaneous island flap from the temporoparietal region pedicled on the parietal branch of the superficial temporalis artery. We obtained efficient and stable repair of the lesion both from a cosmetic and a functional point of view. We were forced to use this procedure for lack of another choice; however, we think that this could become a useful option in covering substance loss in this area when simpler solutions are not available.     

Fetal fracture healing in a lamb model.

A large animal model to assess fetal fracture repair and the ability to close excisional bony defects is presented. Incisional and excisional ulnar fractures were made in 14 midgestation fetal lambs, harvested at serial time points, and subjected to high-resolution low-kilovolt magnification radiographs, magnetic resonance imaging scans, and histologic analysis. Fetal fracture healing was characterized by early closure of excisional defects and rapid fracture healing with minimal or no soft-tissue inflammation or callus formation. Magnetic resonance imaging scans of the fractures revealed a characteristic pattern compatible with the histologic findings, namely, minimal inflammation in soft tissue adjacent to the fracture site. Histologic and magnification radiographic findings indicated that complete bony repair occurred within 21 days in incisional defects and within 40 days in excisional defects. In both cases, healed fetal bone resembled normal bone matrix. Excisional defects, including periosteum, of greater than three times the width of the bony cortex closed rapidly with virtually normal-appearing bony matrix and with minimal or no callus formation.     

Identification of rabbit eosinophils and heterophils in cutaneous healing wounds

Summary The study of wound healing has traditionally used the rabbit as an experimental model. We have recently localized the production of the multifunctional cytokine, TGF-, to eosinophils in rabbit skin wounds. It was evident that during the process of TGF- localization, the distinction between the two granulocytic cell types, eosinophils and heterophils, was impossible by conventional histochemical techniques. This paper describes a rapid method to distinguish these two granulocytes by virtue of their endogenous peroxidases and differential resistance to blockade by inhibitors. In sections that have been blocked by hydrogen peroxide, the peroxidase substrate 3,3-diaminobenzidine, together with nickel chloride (DAB-Ni), preferentially stained the cytoplasm of rabbit eosinophils while sparing those of heterophils. This selective DAB staining of rabbit eosinophil peroxidase in H₂O₂-blocked rabbit wounds was verified at the ultrastructural level by electron microscopy. We applied this technique to quantify eosinophil and heterophil infiltration into the 21-day rabbit cutaneous healing wound model. Heterophils were found infiltrated into all three layers of the wound (clot > granulation > base), but eventually all disappeared by day 21. As with the heterophils, eosinophils which had infiltrated into the clot and base of the wound had disappeared by day 21. Unlike the heterophils, eosinophils in the granulation layer of the wound continued to increase up to day 21. The continually increased and sustained presence of the eosinophils together with their demonstrated production of TGF-, in the granulation layer of the healing wound suggests that these cells play an important role in the organizational aspects of healing wounds.     

A model for the study of epithelial migration in wound healing.

A model is described which enables the detailed study of epithelial regeneration in experimentally produced lesions in the common bile duct of the rabbit. The circular on slightly oval defect of 1 mm diameter produced by a specially developed apparatus has a perfectly smooth base. Epithelial migration in this model has been investigated using light microscopy of transverse sections and scanning electron microscopy of whole preparations. Typical changes in the border cells, characterised by the formation of tapered protusions, can be observed as early as two hours after the lesion has been made. Later the cells in the flattened edge of the moving border also show various types of protrusion which rest on the substratum. Mitotic activity in the surface epithelium and crypts in the surrounding region only increases after closure of the lesion, which usually takes place within 16--24 h.     

Differential expression of transforming growth factor-beta receptors in a rabbit zone II flexor tendon wound healing model

Flexor tendon repair in zone II is complicated by adhesions that impair normal postoperative gliding. Transforming growth factor-beta (TGF-beta) is a family of growth factors that has been implicated in scar formation. The TGF-beta family of proteins binds to three distinct classes of membrane receptors, termed RI, RII, and RIII. In this study, we analyzed the temporal and spatial distribution of TGF-beta receptor isoforms (RI, RII, and RIII) in a rabbit zone II flexor tendon wound healing model.Twenty-eight adult New Zealand White rabbit forepaws underwent isolation of the middle digit flexor digitorum profundus tendon in zone II. The tendons underwent transection in zone II and immediate repair. The tendons were harvested at increasing time points: 1, 3, 7, 14, 28, and 56 days postoperatively (n = 4 at each time point). The control flexor tendons were harvested without transection and repair (n = 4). Immunohistochemical analysis was used to detect the expression patterns for TGF-beta receptors RI, RII, and RIII.Immunohistochemical staining of the transected and repaired tendons demonstrated up-regulation of TGF-beta RI, RII, and RIII protein levels. TGF-beta receptor production in the experimental group (transection and repair) was concentrated in the epitenon and along the repair site. Furthermore, the TGF-beta receptor expression levels peaked at day 14 and decreased by day 56 postoperatively. In contrast, minimal receptor expression was observed in the untransected and unrepaired control tendons.These data provide evidence that (1) TGF-beta receptors are up-regulated after injury and repair; (2) peak levels of TGF-beta receptor expression occurred at day 14 and decreased by day 56 after wounding and repair; and (3) both the tendon sheath and epitenon have the highest receptor expression, and both may play critical roles in flexor tendon wound healing. Understanding the up-regulation of TGF-beta isoforms and the up-regulation of their corresponding receptors during flexor tendon wound healing provides new targets for biomolecular modulation of postoperative scar formation.     

Perforator-based flap in rats: a new experimental model.

A new type of flap, the perforator-based flap, has been described in the last decade. It has been used successfully as a pedicle or free flap by many plastic surgeons. There is no animal model for research, although these flaps have gained popularity in clinical use. We created a perforator-based flap model in the rat (a perforator-based flap group and two control groups; 10 rats in each group) and evaluated the survival characteristics of the new flap. The abdominal skin flap was elevated based on the second perforator of the right superior deep epigastric artery and then sutured to its original bed. In the first control group, the same flap was elevated with a subcutaneous pedicle without any perforator; in the second control group, a right-sided, random-pattern pedicle abdominal skin flap with the same dimensions and location was elevated and sutured to its original bed. Flap survival was studied, and microangiography and histologic studies were performed. The amount of viable skin in the three groups was compared 1 week later. The area of surviving skin paddles in the experimental group ranged from 74 to 83 percent; in the first control group, it was 0 percent; and in the second control group, it ranged from 29 to 44 percent (p < 0.001 and p < 0.001, respectively). There was a predictable and constant area of necrosis in the model.The results of this study demonstrate that most of the abdominal skin of the rat can survive on the basis of a single musculocutaneous perforator vessel. This flap can be easily elevated, and it can be used as a reliable model for flap research.     

Serologic and mucosal immune response to rotavirus infection in the rabbit model.

We examined the humoral immune response to rotavirus infection in specific pathogen-free rabbits inoculated and challenged orally with rabbit Ala rotavirus (7.5 x 10(5) to 1 x 10(7) PFU). The humoral immune response in both serologic and mucosal samples was monitored by using total antibody enzyme-linked immunosorbent assays (ELISAs), isotype-specific ELISAs, and plaque reduction neutralization assays. Following a primary infection, all rabbits shed virus and serologic and mucosal antibody responses were initially detected by 1 week postinoculation. Intestinal immunoglobulin M was detected by 3 days postinoculation, and secretory immunoglobulin A was detected by 6 days postinoculation. Following challenge, rabbits were protected (no detectable virus shedding) from infection. An anamnestic immune response was observed only with mucosal neutralizing antibodies, and all serologic and mucosal immune responses persisted at high levels until at least 175 days postchallenge (204 days postinoculation). Detection of neutralization responses was influenced by the virus strain used in the neutralization assay; all inoculated rabbits developed detectable serum and intestinal neutralizing antibodies against the infecting (Ala) virus strain. Neutralization activity in both serum and mucosal samples was generally, but not exclusively, homotypic (VP7 serotype 3) after both primary and challenge inoculations with Ala virus. Heterotypic serum neutralization activity was observed with serotype 8 (9 of 12 rabbits) and 9 (12 of 12 rabbits) viruses and may be based on reactivity with the outer capsid protein VP4 or on a shared epitope in the C region of VP7. Comparisons of heterologous (serotype 3) and heterotypic neutralizing responses in mucosal and serologic samples revealed that 43% (21 of 49) of the responses were discordant. In 19 of 49 (39%) of these cases, a heterotypic serologic response was seen in the absence of a heterotypic mucosal response, but in 2 of 49 (4%) instances, a heterotypic mucosal response was seen in the absence of a concomitant serologic response. These results provide insight into factors which may affect detection of heterotypic responses.     

Skin recycling following neovascularization using the rat musculocutaneous flap model.

The recycling of a skin territory as part of a musculocutaneous flap despite prior division of existing musculocutaneous perforators or vice versa within an axial cutaneous flap using skin from a previous musculocutaneous flap may sometimes be done safely if an adequate time period has been allotted to permit sufficient neovascularization from adjacent tissues. In order to test this clinically observed phenomenon, a musculocutaneous flap model based on perforators from the rat rectus abdominis muscle was developed and observed to have complete reliability. Groups of five Sprague-Dawley rats each were sequentially utilized to prove that by a single week following creation of a rectus abdominis musculocutaneous flap adequate peripheral neovascularization would evolve to permit total viability of secondary axial epigastric cutaneous flaps incorporating the same skin that initially was the cutaneous portion of the muscle flap. The converse was also confirmed possible, again using sequential groups of five rats each, in that by 2 weeks the skin of an initial abdominal cutaneous flap could instead be safely transposed and nourished as part of a rectus abdominis musculocutaneous flap. The proposition concerning the reliable reuse of identical skin territories as part of disparate metachronous flap configurations appears to be valid.     

Renal transplantation in the rabbit: a model for preservation studies.

Transplantation is necessary for evaluation of kidney preservation procedures, and a model using a small laboratory animal is desirable. The rabbit was found to be a suitable animal for this purpose. Even long periods of anaesthesia without artificial respiration were safely achieved. Hydration and serum electrolytes could be maintained within normal ranges with intravenous injections of isotonic saline and dextrose during and after the operation. The kidneys were implanted by anastomosing the artery and vein end-to-side to the abdominal aorta and the posterior vena cava respectively. The ureter was implanted into the bladder over a nylon stent. In a recent 100 transplantations the incidence of vascular thrombosis was low (4%), but rather more (10%) mainly late ureteral complications were encountered. Transplanted kidneys showed good function with mean peak serum creatinines of 285 mumol/l and normal macroscopic and histological appearance at autopsy.     

The influence of indomethacin on collagen synthesis during tendon healing in the rabbit

The influence of indomethacin on collagen synthesis in intact and healing plantaris longus tendons in the rabbit was investigated. Forty-four male New Zealand White rabbits were subjected to a standardized trauma (tenetomy + repair) on the left hindlimb. Half of the animals were subsequently treated with indomethacin, 10 mg/kg per day orally, and the other half with placebo. After 2 and 4 weeks the rabbits were injected intravenously with 3H-proline and killed 18 h later. Indomethacin affected the collagen metabolism differently depending on whether the tendons were involved in wound healing or not. In intact tendons the drug caused a small general inhibition of collagen synthesis. In the healing tendon there was a shift towards the synthesis of more insoluble collagen with little effect on the total synthesis. After 4 weeks there was also a slight but significant decrease in the amount of hydroxyproline in the most soluble collagen fraction from the tenotomized, indomethacin treated tendons.