IMPORTANCE: Airway transplantation could be an option for patients with proximal lung tumor or with end-stage tracheobronchial disease. New methods for airway transplantation remain highly ...controversial. OBJECTIVE: To establish the feasibility of airway bioengineering using a technique based on the implantation of stented aortic matrices. DESIGN, SETTING, AND PARTICIPANTS: Uncontrolled single-center cohort study including 20 patients with end-stage tracheal lesions or with proximal lung tumors requiring a pneumonectomy. The study was conducted in Paris, France, from October 2009 through February 2017; final follow-up for all patients occurred on November 2, 2017. EXPOSURES: Radical resection of the lesions was performed using standard surgical techniques. After resection, airway reconstruction was performed using a human cryopreserved (−80°C) aortic allograft, which was not matched by the ABO and leukocyte antigen systems. To prevent airway collapse, a custom-made stent was inserted into the allograft. In patients with proximal lung tumors, the lung-sparing intervention of bronchial transplantation was used. MAIN OUTCOMES AND MEASURES: The primary outcome was 90-day mortality. The secondary outcome was 90-day morbidity. RESULTS: Twenty patients were included in the study (mean age, 54.9 years; age range, 24-79 years; 13 men 65%). Thirteen patients underwent tracheal (n = 5), bronchial (n = 7), or carinal (n = 1) transplantation. Airway transplantation was not performed in 7 patients for the following reasons: medical contraindication (n = 1), unavoidable pneumonectomy (n = 1), exploratory thoracotomy only (n = 2), and a lobectomy or bilobectomy was possible (n = 3). Among the 20 patients initially included, the overall 90-day mortality rate was 5% (1 patient underwent a carinal transplantation and died). No mortality at 90 days was observed among patients who underwent tracheal or bronchial reconstruction. Among the 13 patients who underwent airway transplantation, major 90-day morbidity events occurred in 4 (30.8%) and included laryngeal edema, acute lung edema, acute respiratory distress syndrome, and atrial fibrillation. There was no adverse event directly related to the surgical technique. Stent removal was performed at a postoperative mean of 18.2 months. At a median follow-up of 3 years 11 months, 10 of the 13 patients (76.9%) were alive. Of these 10 patients, 8 (80%) breathed normally through newly formed airways after stent removal. Regeneration of epithelium and de novo generation of cartilage were observed within aortic matrices from recipient cells. CONCLUSIONS AND RELEVANCE: In this uncontrolled study, airway bioengineering using stented aortic matrices demonstrated feasibility for complex tracheal and bronchial reconstruction. Further research is needed to assess efficacy and safety. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT01331863
Background Seven years of experimental research provided a valuable tracheal substitute, the aortic allograft, which can promote the regeneration of epithelium and cartilage. In human application, ...both fresh and preserved aortic allografts could be used. The optimal method of aortic allograft preservation remains to be evaluated. This study assessed the use of cryopreserved, decellularized, or glutaraldehyde-treated aortic allografts as tracheal substitutes. Methods Twenty-two sheep underwent tracheal replacement using cryopreserved (n = 10), decellularized (n = 7) or glutaraldehyde-treated (n = 5) allografts, supported by a temporary stent to prevent airway collapse. Aortic segments were retrieved at regular intervals up to 12 months after implantation to analyze the regenerative process. Results All animals survived the operation. Major complications such as infection, stent migration, or obstruction were predominantly encountered in the decellularized group. The lack of major inflammatory response within the aortic graft observed in the glutaraldehyde group was associated with the absence of tracheal regeneration. Histologic examinations showed a progressive transformation of the aorta into a tracheal tissue comprising respiratory epithelium and cartilage only in the cryopreserved group. Conclusions This study demonstrated that regeneration of a functional tissue could be obtained after tracheal replacement with a cryopreserved aortic allograft. The regenerative process followed the same pattern as previously described for fresh allografts. Cryopreserved aortic allografts present major advantages: availability in tissue banks, permanent storage, and no need for immunosuppression. This offers a new field of perspectives for clinical application in patients with extensive tracheal cancer.
Bronchial Replacement With Arterial Allografts Radu, Dana M., MD; Seguin, Agathe, MD; Bruneval, Patrick, MD, PhD ...
The Annals of thoracic surgery,
07/2010, Volume:
90, Issue:
1
Journal Article
Peer reviewed
Open access
Background Pneumonectomy is well known for a high risk of postoperative death. The alternative, sleeve lobectomy, is sometimes technically inaccessible, and is associated with locoregional ...recurrence. In certain situations, the use of a bronchial substitute would allow longer bronchial resections with better security margins. Previous experiments demonstrated that aortic grafts are valuable tracheal and carinal substitutes. The present study evaluated bronchial replacement with arterial allografts. Methods Fifteen female sheep underwent a left bilobectomy with replacement of the bronchus intermedius with arterial allografts: 5 received a fresh graft (group 1) and 10 received cryopreserved (group 2). A bronchial silicone stent was used to confer rigidity. Evaluation was conducted on clinical and histologic criteria at regular intervals up to 18 months. Results There were no perioperative deaths. Atelectasis, the only early postoperative complication (n = 2), was successfully treated by fiberscopic aspiration. The late postoperative period was uneventful in 12 sheep. Complications included 1 bronchopneumonia, 1 pulmonary abscess, and 1 distortion of the bronchial stent. Fiberscopic examination revealed 3 sheep with granuloma formation. The bronchial stent was removed in 3 sheep, 1 at 9 months and 2 at 12 months, without clinical complications or stenosis of the graft. Histologic analysis showed regeneration of new bronchial tissue, comprising epithelium and cartilage. Conclusions This study confirmed that an arterial allograft could be a valuable bronchial substitute. The use of a bronchial substitute offers new perspectives in surgical resection of lung cancer because it would avoid pneumonectomy in some patients.
Background Pneumonectomies for lung cancer are associated with a high postoperative mortality, especially when right-sided, after neoadjuvant radiochemotherapy, and in patients over 70 years of age. ...Preliminary studies in our laboratory have shown that aortic grafts could be valuable airway substitutes. We report the first human bronchial transplantation of a cryopreserved aortic allograft used as a biologic airway substitute to prevent a pneumonectomy for lung cancer. Methods The procedure was performed in a high-risk 78-year old patient with an extensive right bronchopulmonary malignant tumor pretreated with chemotherapy. After a complete resection of the lung cancer using an upper bilobectomy with lymph node removal, mobilization procedures did not allow for a primary end-to-end bronchial anastomosis. A stent-supported cryopreserved aortic allograft from a certified tissue bank was interposed to restore the bronchial continuity with sparing of the lower lobe. Results The postoperative course was eventful for a supraventricular arrhythmia leading to mild pulmonary edema that resolved using standard medical therapy, and a right lower lobe atelectasis with bacterial colonization that required fiberoptic bronchoscopies in addition to antibiotic treatment. A 1-year postoperative evaluation found a well-functioning reimplanted lower lobe with no complications related to the cryopreserved aortic allograft or the stent. The patient recovered to his baseline activity with a satisfying health-related quality of life. Conclusions We demonstrate the feasibility of this surgical innovation to prevent the high-risk procedure of pneumonectomy in a single case. If confirmed in larger series of selected patients, it could bring new perspectives in conservative lung cancer surgery.
BACKGROUND: Weak D Types 1, 2, and 3 recipients cannot be immunized when exposed to D antigen. Molecular biology is very efficient to type weak D variants but rarely implemented in daily practice. ...The serologic typing practice of weak D in a Caucasian patient population was analyzed and a transfusion strategy is proposed.
STUDY DESIGN AND METHODS: Samples typed either ddCcee or ddccEe in routine laboratories were tested with the indirect antiglobulin test (Du test). Du‐positive samples were screened for weak D alleles Types 1, 2, and 3 and further tested with immunoglobulin M (IgM) anti‐D reagents, used in a fully automated device.
RESULTS: A total of 468 of 55,162 samples were found to be ddCcee or ddccEe. Ninety‐three expressed weak D after the Du test leading to D+ assignment for transfusion. Seventy‐three percent of Du‐positive samples were weak D alleles Type 1, 2, or 3. Almost all weak D Types 1, 2, and 3 were positive with IgM reagents in gel matrix with an automated device. Other variants that could be potentially associated with anti‐D alloimmunization, however, were also positive.
CONCLUSION: Serology is very sensitive to detect weak D Types 1, 2, and 3, but there is no cutoff to distinguish variants of clinical significance. When molecular analysis is not available, it is proposed that a D+ status for blood recipients found to be weak D with a sensitive method be assigned, except for women of childbearing age or younger, because of the remaining possibility to be partial D or other rare weak D who can be immunized.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
Abstract A new protocol for cryopreservation of arteries frozen at −80 °C was compared to the reference protocol for cryopreservation at −150 °C and to freshly harvested arteries. The aim of the ...study is to evaluate both protocols as global procedures to freeze and thaw arteries commonly used in tissue banks. Changes in mechanical properties of rabbit common carotid arteries were studied. Vascular segments were tested in vitro under dynamics loading conditions. Pressure and diameter were recorded simultaneously by a high fidelity transducer and an echotracking device, respectively. The pressure–diameter relationship was fitted by the arctangent Langewouters’ model and the arterial thickness was derived from histological measurements. Histological sections showed that the fresh and −80 °C groups were less damaged by hemodynamic load and histological preparation than the −150 °C group ( p < 0.05). No differences between fresh and cryopreserved arteries regarding the structural (diameter, intimal-media thickness) and mechanical parameters (distensibility, circumferential stress, elastic modulus) were found. The isobaric circumferential stress was reduced in frozen arteries. These results demonstrate that the cryopreservation at −80 °C preserves the histological structure and mechanical properties better than the cryopreservation at −150 °C, suggesting that the new cryopreservation protocol at −80 °C is a method of choice for treating vessel replacement in vascular surgery.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
7.
Mesure de l’hématocrite des produits de thérapie cellulaire Chartois, Anne-Gaele; Simon, Philippe; Fialaire-Legendre, Anne ...
Transfusion clinique et biologique : journal de la Société française de transfusion sanguine,
September 2017, 2017-09-00, Volume:
24, Issue:
3
Journal Article
Peer reviewed
Les greffes allogéniques se font souvent dans un contexte d’incompatibilité ABO majeure, nécessitant le calcul du volume de globules rouges (GR) à partir de l’hématocrite or l’hyperleucocytose des ...greffons peut être responsable d’une surestimation de l’Ht.
Une étude multicentrique réalisée sur des prélèvements de CSH compare les résultats d’Ht obtenus sur des automates d’hématologie (AH) ainsi que les Ht calculés à partir des GR ou de l’hémoglobine(Hb). L’effet «̊matrice̊» sur la mesure de l’Ht a été également évaluée.
Les paramètres sanguins d’échantillons de CSH périphériques ont été mesurés sur différents AH par des unités de thérapie cellulaire. L’impact de la matrice hyperleucocytaire a été évaluée sur une suspension de GB préparés à partir de couches leucoplaquettaires additionnées de 5 % de GR. La mesure de l’Ht a été réalisée sur 3 AHC et par méthode manuelle.
Les hématocrites diffèrent suivant la technologie des AH. Le biais dose̊–̊dépendante augmente avec la concentration croissante des GB dans les échantillons. Les comparaisons entre les Ht mesurés sur l’AH de référence et les Ht calculés à partir du nombre des GR (Ht=GR×9) ou de l’Hb (Ht=Hb×3) montrent une diminution du biais. Par ailleurs, on constate une augmentation progressive de l’Ht sur la «̊matrice̊» diluée+5 % de GR.
On observe une surestimation variable de l’Ht suivant les AH plus marquée sur les produits très hyperleucocytaires. Les Ht mesurés sur les AH combinant impédance et focalisation hydrodynamique sont les plus faibles. L’étude permet de faire un état des lieux des techniques et de leur performance au regard de la détermination de l’Ht dabs les PTC.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK, ZRSKP
Selon la monographie 2.6.27 de la pharmacopée européenne (contrôle microbiologique des produits cellulaires), la comparaison interlaboratoires est une possibilité de validation de méthodes et la ...participation à des études collaboratives permet à chaque laboratoire d’évaluer ses performances et de répondre à ces exigences de validation.
En 2016, en relais de l’ANSM, le laboratoire de contrôle qualité de l’EFS Aquitaine-Limousin a organisé deux campagnes de comparaison inter-laboratoires, l’un portant sur le contrôle microbiologique des milieux cornéens, l’autre sur celui des produits de thérapie cellulaire.
Le matériel à contaminer était dans le premier cas un milieu de transport de cornée, dans le deuxième cas des résidus de couches leuco-plaquettaires du sang total. Les germes contaminants ont été choisis parmi les germes recommandés pour la validation de méthode ou pour leur intérêt. Les laboratoires participants à ces études utilisent des techniques différentes (automatisée ou manuelle), des durées et des températures d’incubation différentes, toutes en accord avec les recommandations de la pharmacopée 2.6.27.
Le contrôle des milieux de cornées a rassemblé 7 participants̊; 86 % des laboratoires ont eu 100 % de réponses satisfaisantes̊: détection d’un échantillon stérile et identification correcte des deux échantillons contaminés.
Dix-sept laboratoires ont participé au contrôle des produits de thérapie cellulaires. Quatre-vingt-deux pour cent des laboratoires ont eu 100 % de réponses satisfaisantes, les erreurs mises en évidence dans les autres laboratoires correspondent à une identification erronée des échantillons ou à une contamination croisée.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK, ZRSKP
Bone allografts are commonly used by orthopedists to provide a mechanical support and template for cellular colonization and tissue repair. There is an increasing demand for bone graft substitutes ...that are safe and easy to store but which are equally effective in supporting new bone growth. In this study, we compared three different human bone allografts: (1) the cryopreserved allograft (frozen), (2) the gamma-irradiated and cryopreserved allograft (γ-irradiated), and (3) the solvent dehydrated and γ-irradiated-processed bone allograft (Tutoplast(®) Process Bone TPB). Human mesenchymal stromal cells (hMSCs) have the potential to differentiate into osteogenic, chondrogenic, and adipogenic lineages. Our results showed that hMSC seeding efficiency was equivalent among the three bone allografts. However, differences were observed in terms of cell metabolism (viability), osteoblastic gene expression, and in vivo bone formation. Frozen allografts had the higher frequency of new bone formation in vivo (89%). Compared with frozen allografts, we demonstrated that TPB allografts allowed optimal hMSC viability, osteoblastic differentiation, and bone formation to occur in vivo (72%). Further, the frequency of successful bone formation was higher than that obtained with the γ-irradiated allograft (55%). Moreover, after hMSC osteoinduction, 100% of the TPB and frozen allografts formed bone in vivo whereas only 61% of the γ-irradiated allografts did. As healthcare teams around the world require bone-grafting scaffolds that are safe and easy to store, the TPB allograft appears to be a good compromise between efficient bone formation in vivo and convenient storage at room temperature.
Stem cells may represent an excellent strategy to improve the healing of skin ulcers. Today the administration mode of stem cells to skin defects remains unsatisfactory. Delivering stem cells with ...topical treatments represents a new strategy and answering the patients' need. Mesenchymal stromal cells (MSC) have been shown to improve wound healing of cutaneous lesions and amniotic membrane (AM) is known to represent a natural scaffold for cells. The aim of this study is to develop a tissue-engineered product combining MSC and AM for clinical use. In this work we investigated whether the stromal matrix of intact human AM could constitute a scaffold for human MSC derived from either bone marrow (BM) or adipose tissue (AT). For this purpose, clinical-grade AM, MSC, and culture medium were used. We performed experiments of short-term adherence and proliferation for 15 days after the seeding of the cells. Morphological aspects and secretion profiles of MSC onto AM were studied, respectively, by scanning electron microscopy and Luminex analysis. Results demonstrated that the stromal matrix allow the adherence in much greater amount of MSC from BM or AT compared to 2D material. Experiments of proliferation showed that both kinds of MSC could proliferate on the stromal matrix and remain viable 15 days after the seeding of the cells. The 3D analysis of MSC culture demonstrated that both types of MSC invaded the stromal matrix and grew in multiple layers while retaining their fibroblastic morphology. By studying the secretion profile of MSC onto the stromal matrix, we found that both kinds of MSC secrete important cytokines and growth factors for wound healing of cutaneous lesions, such as vascular endothelial growth factor, hepatocyte growth factor, and basic fibroblast growth factor. In conclusion, these results suggest that the stromal matrix of AM seeded with MSC represents a bioactive scaffold that should be evaluated in patients with a nonhealing cutaneous wound.
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