Xenogenic transplantation of human breast adipose-derived stromal vascular fraction enhances recovery of erectile function in diabetic mice.

The adipose tissue-derived stromal vascular fraction (SVF) is an ideal source of stem and stromal cells. The aim of this study was to examine whether and how xenogenic transplantation of human breast SVF restores erectile function in diabetic mice. Human SVF was isolated from five patients (age, 20-45 yr) undergoing reduction mammoplasty. Eight-week-old C57BL/6J mice were used, and diabetes was induced by intraperitoneal injection of streptozotocin. At 8 wk after induction of diabetes, the animals were randomly distributed into controls and diabetic mice treated with a single intracavernous injection of PBS, human SVF at different concentrations, or human SVF lysate. Two weeks later, erectile function was measured by cavernous nerve stimulation, and the penis was then harvested for biochemical examinations. Erectile function was significantly improved in diabetic mice treated with human SVF (2 × 10(5), 5 × 10(5), and 1 × 10(6) cells/20 µl) and SVF lysate. Human SVF treatment in diabetic mice significantly increased cavernous endothelial and smooth muscle cell contents, induced eNOS phosphorylation, and restored penile nNOS-positive nerve fibers. Human SVF lysate induced secretion of angiogenic factors and expression of their receptors. Human SVF did not increase serum levels of proinflammatory cytokines. A limitation of this study was that the exact composition of the human SVF was not examined. In summary, xenogenic transplantation of human SVF did not induce systemic inflammation and successfully improved erectile function in diabetic mice through enhanced penile angiogenesis and neural regeneration.

Breast reconstruction with autologous fat graft mixed with platelet-rich plasma.

BACKGROUND: The purpose of this study was to review the authors' experience of fat grafting, evaluating the effects related to the use of fat grafting with platelet-rich plasma (PRP) in the improvement of fat volume in breast reconstruction and comparing the results with a control group (only centrifuged fat grafting). METHODS: A total of 50 patients aged between 19 and 60 years affected by breast soft-tissue defects were analyzed at the Plastic and Reconstructive Department of the University of Tor Vergata. They were treated with fat grafting + PRP. The control group (50 patients with breast soft-tissue defects) were treated with centrifuged fat grafting injection according to Coleman's procedure. RESULTS: The patients treated with PRP added to the autologous fat grafts showed a 69% maintenance of the contour restoring and of 3-dimensional volume after 1 year, whereas the patients of the control group treated with centrifuged fat grafting showed a 39% maintenance. CONCLUSION: PRP mixed with fat grafting leads to an improvement in maintaining breast volume in patients affected by breast soft-tissue defects.

High and low mammographic density human breast tissues maintain histological differential in murine tissue engineering chambers.

Mammographic density (MD) is the area of breast tissue that appears radiologically white on mammography. Although high MD is a strong risk factor for breast cancer, independent of BRCA1/2 mutation status, the molecular basis of high MD and its associated breast cancer risk is poorly understood. MD studies will benefit from an animal model, where hormonal, gene and drug perturbations on MD can be measured in a preclinical context. High and low MD tissues were selectively sampled by stereotactic biopsy from operative specimens of high-risk women undergoing prophylactic mastectomy. The high and low MD tissues were transferred into separate vascularised biochambers in the groins of SCID mice. Chamber material was harvested after 6 weeks for histological analyses and immunohistochemistry for cytokeratins, vimentin and a human-specific mitochondrial antigen. Within-individual analysis was performed in replicate mice, eliminating confounding by age, body mass index and process-related factors, and comparisons were made to the parental human tissue. Maintenance of differential MD post-propagation was assessed radiographically. Immunohistochemical staining confirmed the preservation of human glandular and stromal components in the murine biochambers, with maintenance of radiographic MD differential. Propagated high MD regions had higher stromal (p = 0.0002) and lower adipose (p = 0.0006) composition, reflecting the findings in the original human breast tissue, although glands appeared small and non-complex in both high and low MD groups. No significant differences were observed in glandular area (p = 0.4) or count (p = 0.4) between high and low MD biochamber tissues. Human mammary glandular and stromal tissues were viably maintained in murine biochambers, with preservation of differential radiographic density and histological features. Our study provides a murine model for future studies into the biomolecular basis of MD as a risk factor for breast cancer.

Round Form-Stable Breast Implants: Diagnosis and Management of Complications.

Silicone gel breast implants have been used for breast augmentation and reconstruction since 1962. Since then, multiple generations of implants have been created in an effort to improve safety and efficacy. Before 1990, silicone gel implants were characterized as having thin shells and non-cohesive gel; however, since then devices are created with cohesive silicone gel and a variety of surfaces. Despite improvements, no implant will last forever; however, these devices are safe and effective based on numerous clinical and epidemiologic studies. As with all medical devices, complications using round form-stable implants for breast reconstruction can occur and will be reviewed in this article.

Validated Outcomes in the Grafting of Autologous Fat to the Breast: The VOGUE Study. Development of a Core Outcome Set for Research and Audit.

BACKGROUND: Autologous fat grafting is an important part of the reconstructive surgeon's toolbox when treating women affected by breast cancer and subsequent tumor extirpation. The debate over safety and efficacy of autologous fat grafting continues within the literature. However, work performed by the authors' group has shown significant heterogeneity in outcome reporting. Core outcome sets have been shown to reduce heterogeneity in outcome reporting. The authors' goal was to develop a core outcome set for autologous fat grafting in breast reconstruction. METHODS: The authors published their protocol a priori. A Delphi consensus exercise among key stakeholders was conducted using a list of outcomes generated from their previous work. These outcomes were divided into six domains: oncologic, clinical, aesthetic and functional, patient-reported, process, and radiologic. RESULTS: In the first round, 55 of 78 participants (71 percent) completed the Delphi consensus exercise. Consensus was reached on nine of the 13 outcomes. The clarity of the results and lack of additional suggested outcomes deemed further rounds to be unnecessary. CONCLUSIONS: The VOGUE Study has led to the development of a much-needed core outcome set in the active research front and clinical area of autologous fat grafting. The authors hope that clinicians will use this core outcome set to audit their practice, and that researchers will implement these outcomes in their study design and reporting of autologous fat grafting outcomes. The authors encourage journals and surgical societies to endorse and encourage use of this core outcome set to help refine the scientific quality of the debate, the discourse, and the literature. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, V.

Superior Pedicle Mammaplasty without Parenchymal Incisions after Massive Weight Loss.

BACKGROUND: After considerable weight loss, the breast suffers significant deformation. The ptotic breast is characterized by a lack of superior pole, tissue excess in the inferior pole, down-migration of nipple-areola complex (NAC) with redundancy of skin tissue. The authors describe a mastopexy technique based on a modulated and progressive reshaping, back rotation, and suspension of mammary gland parenchyma without parenchymal incisions. METHODS: Forty-five patients with bilateral moderate or severe breast ptosis underwent mastopexy from January 2011 to January 2014 with complete detachment of breast from the pectoralis major muscle and the plication of parenchyma without any parenchymal incision. Patients were followed up for one year, reporting any complication, and measuring the jugulum-NAC distance. The outcomes were assessed by the patients as well as the surgical team. RESULTS: The aesthetic outcomes were good or excellent in all patients. The new mammary contour and the distance between the jugular fossa and the nipple were stable during this time with a good filling of upper pole. No major complications were reported. CONCLUSIONS: This technique gave good breast shape, long-term projection, and upper pole fullness, without parenchymal incisions. It restores breast shape and projection, especially in post-bariatric patients. A similar technique has not been described yet.

Growth of human normal and neoplastic mammary tissues in the cleared mammary fat pad of the nude mouse.

Dysplastic and malignant human breast tissues were grown successfully in the cleared mammary fat pads (CFP) of nude mice. The mammary fat pads were cleared while the mice were in a germfree isolator. Prepared mice were removed fron the germfree enviornment to facilitate transplantation of the human mammary tissue into their CFP and subsequently were maintained in sterile laminar flow racks.

Long-term explant culture of normal mammary epithelium.

Mammary epithelium and surrounding stroma have been maintained in an explant system for 1 to 6 months and subsequently xenografted into athymic nude mice. The morphological characteristics of 26 cases of normal human mammary epithelium in long-term explant culture were described, using high-resolution light and electron microscopy. Normal human breast tissue specimens were obtained from immediate autopsy or surgical resection. The explants were cultured in Connaught Medical Research Laboratories Medium 1066 supplemented with serum, insulin, and hydrocortisone. The histotypic features of the mammary epithelium in both the central portion of the explant and the epithelial outgrowth onto the surface of the explant were described. In some cultures, the cells acquired more keratin and formed multicellular blister-like domes. Tissues from eight cases after 1 to 14 weeks in culture were xenografted in athymic nude mice and were maintained up to 924 days.

Preneoplastic lesions of the human mammary gland transplanted into the nude athymic mouse.

Morphologically normal lobules and atypical lobules postulated precancerous to ductal carcinoma were transplanted to test their biological behavior. Supravital staining disclosed the 1 to 4-mm microorgans. "Cleared" mammary fat pads of nude mice were optimal transplantation sites. Of the total of 217 transplants from 19 cancer-associated and 13 non-cancer-associated breast, 151 survived after 2 to 27 weeks. Of 61 surviving normal-appearing lobules from cancer-associated breasts transplanted without prior in vitro maintenance, 20 (30%) dedifferentiated, and of 48 surviving lobules from noncancerous breasts, 11 (20%) dedifferentiated. Fifteen of 28 histologically normal-appearing lobules (60%) obtained from cancer-associated breasts after age 50 dedifferentiated. Thirty of 36 atypical lobules isolated from the breast tissue were obtained from cancer-associated breasts and the 5 of those that dedifferentiated came from cancer-associated breasts. Twenty of 22 (90%) dedifferentiating transplants from cancer-associated breasts showed a vascularization response, whereas 3 of 7 (43%) from noncancer-associated breasts did so. If dedifferentiation in this experimental setting is indicative of a precancerous potential, the data on normal-appearing lobules obtained from cancer-associated breasts from women over age 50 suggest that these lobules carry the greatest precancerous potential. Such lobules probably belong to a type persisting after menopause and they may be hormonally autonomous. Such lobules might undergo further atypia in vivo and, eventually, cancerous transformation.

Breast sharing: New perspectives on an old method.

BACKGROUND: Many techniques have been described for breast reconstruction after mastectomy throughout the decades. In many cases, with excess tissue being discarded, a significant reduction of the contralateral breast was needed for symmetry. Described by Marshall as a one-staged, autologous, non-microsurgical breast reconstruction technique, this method was used in perforator flaps as a breast-sharing technique. METHODS: Between June 2011 and January 2014, the breast-sharing technique was performed in seven patients with simple mastectomy, delayed breast reconstruction, and willingness for autologous non-microsurgical breast reconstruction. All the participants in this study received preoperative oncological screening with ultrasound, magnetic resonance imaging, or mammography, which revealed the absence of pathologic imaging in the donor breast. RESULTS: The experiences of seven patients who underwent breast reconstruction surgery through breast-sharing technique are presented. Due to venous congestion, one of the patients (14%) suffered major complications with total loss of the flap. A total of four patients (57%) incurred minor complications with little to no repercussions on the final outcome. The functional and aesthetic outcomes were very satisfactory, and only one patient required a second touch-up surgery for lipofilling due to unsatisfactory breast volume. Regular follow-ups were done by the oncologist with no recurrences found up to the present. CONCLUSIONS: Using contralateral breast as the donor site, this article presents the first case series for one-stage mammarian reconstruction. With some complications but good aesthetic outcomes, this method has been shown as another available method for breast reconstruction in patients with hypertrophic and ptotic breast.

A novel method for growing human breast epithelium in vivo using mouse and human mammary fibroblasts.

A novel system is described for studying the growth of normal human mammary epithelium in vivo as grafts in athymic nude mice. The key feature of this model is reconstitution of the epithelial-stromal interactions required for normal growth and differentiation of the human mammary epithelium, which produces ducts that are comparable to those in the normal human mammary gland. Human breast epithelial organoids were combined with mammary fibroblasts from mouse or human origin in collagen gels, which were subsequently transplanted under the renal capsule of female nude mice hosts. The resulting grafts showed an increase in the ductal density compared with that observed previously. These ducts expressed appropriate markers for luminal and myoepithelial cells and steroid receptors. Treatment of the host with diethylstilbestrol or estradiol and progesterone significantly increased the number of ducts observed and increased cell proliferation. The grafts also displayed production of beta-casein and milk fat globule membrane protein when the hosts were allowed to become pregnant. This model allows for a variety of epithelial and stromal cells to be used in combination, which would aid in understanding key factors that regulate normal human mammary gland development.

Phenotypic characterization of collagen gel embedded primary human breast epithelial cells in athymic nude mice.

We have developed a method to characterize the phenotypes and tumorigenicity of dissociated human breast epithelial cells. The dissociated cells were first embedded in collagen gels and subsequently transplanted subcutaneously in vivo in athymic nude mice. The transplantation of dissociated epithelial cells from reduction mammoplasties, presumed to be normal, always resulted in normal histomorphology. Epithelial cells were arranged as short tubular structures consisting of lumina surrounded by epithelial cells with an occasional more complex branching structure. These outgrowths were surrounded by intact basement membrane and were embedded in collagen gel that, at termination, contained collagenous stroma with fibroblasts and blood vessels. In contrast, transplantation of dissociated breast epithelial cells from breast cancer specimens resulted in outgrowths with an invasive pattern infiltrating the collagen gel as well as frank invasion into vascular space, nerves and muscles. These observations were made long before the subsequent palpable stage which resulted if left in the mouse for a long enough time. The dissociated human breast epithelial cells thus retained their intrinsic property to undergo morphogenesis to reflect their original phenotype when placed in a suitable environment, the collagen gel.

Carcinoma in a transplanted nipple.

A 26-year-old woman requested that the nipple from her breast be saved for later reconstructive mammoplasty. At the time of mastectomy, two biopsy specimens from the base of the nipple showed no malignant neoplasms, The nipple was transplanted to the left thigh. Eleven months later, a 5-cm mass developed deep in the nipple transplant, and an adjacent inguinal lymph node became enlarged. Examination of biopsy specimens from the base of the nipple transplant showed the same duct cell carcinoma as the original specimen.

Estrogen and progesterone receptors can be maintained in normal human breast epithelial cells in primary culture and after transplantation into nude mice.

The estrogen and progesterone receptor expression was determined in breast epithelial cells from reduction mammoplasty specimens. Only a subset of the luminal epithelial cell population was found to be positive for these receptors. When these cells were subsequently cultured in 3-dimensional collagen gel system (in vitro model) or transplanted into nude mice (in vivo model), the cells retained their ability to express both receptors in these experimental systems. The maintenance of primary normal human breast epithelial cells expressing the estrogen and progesterone receptors in experimental systems has not been reported previously.

Montgomery's areolar tubercle. A light microscopic study.

Conflicting impressions regarding the anatomy of Montgomery's areolar tubercle exist. Twelve modified radical mastectomy specimens provided 1,536 serial sections of areolar tubercles. In 34 of 35 tubercles (97%), a mammary lactiferous duct was associated with a sebaceous apparatus. This lactiferous duct ascended from deeper mammary parenchyma and entered the sebaceous gland. Histopathologic changes identified included featured of fibrocystic disease, atypical intraductal hyperplasia, and carcinoma in situ. Because the areolar tubercle has two components, a sebaceous gland and a mammary duct arising from deeper breast parenchyma, diseases of the breast may also involve the areola independent of papilla-nipple involvement. Areolar preservation may best be used with the knowledge that diseases underlying the areola may also involve the areola.

Graft of breast tissue for correction of congenital asymmetry.

This represents a case of a graft of autogenous breast gland from the enlarged side to augment the hypoplastic breast in cases of congenital asymmetry. Long-term consequences of this transfer and the thickness of the transferred gland that could be used remain unknown. It was with the patient's wishes and agreement to close follow-up that we undertook this approach to see if we could find a one-step procedure for correction of asymmetry without the use of either foreign material or flap. Mammogram at 2 years is normal.

[Autotransplantation of nipple-areolar complex in a modified radical mastectomy--indications and three-stage breast reconstruction].

Necessity of breast reconstruction after mastectomy has been increasing in recent years. For better reconstruction, we preserve a nipple-areolar complex (NAC) by transplanting it temporarily onto the lower abdominal wall and retransplant it to the restored breast mound in a subsequent operation. Indications for NAC preservation are as follows: (1) by palpation the tumor is found to be smaller than 3.0cm in diameter without apparent regional and distant metastasis, (2) neither abnormal nipple discharge nor nipple retraction is observed, (3) tumor is remote more than 3.0cm from the areolar margin, (4) no abnormal shadows are seen below the nipple and areola in the mammogram, (5) no microscopic extension of the cancerous cells is detected beneath the resected NAC. We have performed 18 transplantations using this procedure with good cosmetic results. There were no recurrent cases due to NAC preservation. It is concluded that this technique for preserving the NAC by autotransplantation is easy to perform and useful for breast reconstruction because of low risk of recurrence and better cosmetic results.

Mamoplastia de aumento com lipoenxertia autóloga: revisão da literatura e relato de caso / Breast augmentation with autologous fat grafting: literature review and case report

INTRODUÇÃO: O autotransplante do lipoaspirado na mama para fins estéticos e reconstrutivas tem avançado intensamente na técnica e na tecnologia, assim como na aceitação dos médicos e dos pacientes. O autor relata um caso de aumento mamário estético e discute a revisão da literatura, interessando a eficácia, o potencial carcinogênico e o exame de imagem. MÉTODO:Revisão da literatura no Pubmed na língua inglesa e na Revista Brasileira de Cirurgia Plástica, e relato de caso da experiência inicial do autor. RESULTADO: O volume manteve-se estável a partir do segundo mês, e não houve complicações no pós-operatório. Os exames de imagem não apresentaram alterações patológicas. Foram selecionados 24 artigos relacionados. DISCUSSÃO: Dos 24 artigos, só há dois artigos prospectivos não controlados, mas, de maneira geral, não há problemas no diagnóstico nos exames por imagem, não há aumento de potencial cancerígeno, e os resultados são bons nas séries de casos. CONCLUSÃO: O procedimento é reprodutível, seguro e eficaz, consolidando-se como uma indicação no tratamento reparadora da mama e uma opção no aumento estético. Entretanto, uma curva de aprendizado mais longa pode ser necessária, para evitar complicações e atingir bons resultados.

INTRODUCTION: The technique and technology lipoaspirate autotransplantation to the breast with the aim of aesthetic appearance and reconstruction has strongly advanced; further, its acceptance by doctors and patients has also improved. The author reports cosmetic breast augmentation and performed a literature review, focusing on the efficacy, carcinogenic potential, and imaging diagnosis. METHOD:A literature review was performed using English-language articles from the PubMed database and the Brazilian Journal of Plastic Surgery (RBCP); in addition, case series of the initial experience of the author has been described. RESULTS: The volume remained stable from the second month, and there were no postoperative complications. Imaging did not show any pathological alterations. In all, 24-related articles were selected. DISCUSSION: Among the 24 articles, only two prospective non-controlled studies were found, but overall, imaging diagnostic tests did not reveal problems, the carcinogenic potential was not increased, and case series had positive results. CONCLUSION: The procedure is reproducible, safe and effective, and reinforces the use of this technique in breast reconstruction and as an option in cosmetic breast augmentation. However, it may require a longer learning curve to avoid complications and achieve good results.