Pre and post muscle samples were collected (~60 min) after the participants had finished a battery of performance tests, as described previously (Horwath et al., 2020 (link)). Muscle biopsies were obtained from the middle portion of the vastus lateralis muscle after the administration of local anesthesia (2–4 ml of 20 mg ml−1 Carbocaine; Astra Zeneca, Södertälje, Sweden). Using the Weil-Blakesley conchotome technique (Ekblom, 2017 (link)), one muscle sample (~50 mg) was snap frozen in liquid nitrogen for later analysis of the mitochondrial proteins. Another sample was quickly dissected free from visible blood and connective tissue, embedded in OCT-medium and frozen in isopentane cooled by liquid nitrogen. This specimen was later used for the immunohistochemical analyses. A third sample (~10 mg) was immediately placed in ice-cold mitochondrial isolation medium upon removal (100 mM sucrose, 100 mM KCl, 50 mM Tris-HCl, 1 mM KH2PO4, 100 μM EGTA, 0.1% BSA; final pH of 7.4), followed by the mitochondria isolation procedure (described later in the methods section).
Carbocaine
Manufactured by AstraZeneca
Sourced in Sweden, Italy
Carbocaine is a local anesthetic used for pain relief. It contains the active ingredient mepivacaine hydrochloride. Carbocaine is designed to temporarily block nerve signals in the body to prevent the sensation of pain.
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Lab products found in correlation
5 protocols using carbocaine
1
Muscle Sampling and Mitochondrial Isolation
2
Vastus Lateralis Muscle Biopsy
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A skeletal muscle sample was obtained from the middle portion of the vastus lateralis muscle at a depth of 2–3 cm, about one-third of the distance from the upper margin of the patella to the anterior superior iliac spine with the participants resting in a semi recumbent position lying on a bench. After local anesthesia (2–4 ml carbocaine 20 mg ml−1; AstraZeneca, Södertälje, Sweden), an incision (0.5–1 cm) was made through the skin and fascia and a muscle sample (50–100 mg) was obtained with the Weil-Blakesley chonchotome technique. A portion of the sample was snap frozen in liquid nitrogen, while second and third portions were rapidly placed in ice-cold mitochondrial isolation medium and relaxing medium (see section below), followed by mitochondria isolation and muscle fiber permeabilization, respectively, as later described.
3
Surgical Extraction of Mandibular Molar
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Surgery was performed under local anaesthesia. The nerve block was achieved with 3% mepivacaine and the soft-tissue infiltration with 2% mepivacaine and 1 : 100,000 adrenaline (Carbocaine, AstraZeneca, Italy). A mucoperiosteal flap was raised by making an incision distal to the lower second molar along the length of the anterior border of the ascending ramus of the mandible. Ostectomy and tooth sectioning were then performed. After completely extracting the tooth, the socket was revised and the flap was sutured with interrupted synthetic nonabsorbable sutures (4/0 Ethilon™, Ethicon Ltd, UK). The surgery duration was measured from the start of the surgical procedure until completing the last suture. The sutures were removed 1 week later.
4
Comprehensive Oral Hygiene Regimen for Implant Surgery
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Before the surgical treatment, all patients underwent an oral hygiene protocol that consisted of polishing when needed, and supra- and subgingival debridement. One hour prior to surgery, patients received prophylactic antibiotic therapy with 2 g of Amoxicillin (Augmentin, GlaxoSmithKline, London, UK). Immediately before the procedure, they were instructed to rinse with a 0.2% chlorhexidine digluconate solution (Corsodyl, GlaxoSmithKline Consumer Healthcare, Genval, Belgium) for 2 min. All surgical procedures were performed by the same experienced surgeon (L.V.S.). Local anesthesia with 2% mepivacaine 1:100,000 adrenalin (Carbocaine, AstraZeneca, Milan, Italy) was used.
The surgical strategy first provided the placement of implants (TPA, AZ implant, San Lazzaro di Savena, Bologna, Italy) at healed sites and then the extractions of the teeth where immediate implants were planned and at the end the others remaining extractions.
After implant insertion and teeth extraction, the multi-unit abutments were screwed (Figure 6 ), and an impression was taken to prepare a provisional screwed retained prosthesis.
The healing screws were screwed after 6 h, the provisional prosthesis was screwed, and an occlusal check was performed (Figure 7 ).
The surgical strategy first provided the placement of implants (TPA, AZ implant, San Lazzaro di Savena, Bologna, Italy) at healed sites and then the extractions of the teeth where immediate implants were planned and at the end the others remaining extractions.
After implant insertion and teeth extraction, the multi-unit abutments were screwed (
The healing screws were screwed after 6 h, the provisional prosthesis was screwed, and an occlusal check was performed (
5
Muscle Biopsy Sampling Protocol
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Muscle biopsies at Baseline and Exit were collected approximately 60 minutes after the subjects had completed a set of performance tests (jump tests, knee extension torque, running time to exhaustion and Wingate anaerobic power test), as described earlier (30) . Biopsies were taken from the middle portion of the vastus lateralis muscle approximately 15 cm above the patellae (at a depth of 2-3 cm). After administration of local anesthesia (Carbocaine 20 mg ml -1 , AstraZeneca AB, Sweden), a small incision was made to the skin and fascia and samples of 50-100 mg were obtained with a Weil-Blakesley conchotome (14) . After quickly removing visible blood or connective tissue, fiber bundles were oriented perpendicular to the horizontal surface and mounted in O.C.T embedding medium (Tissue-Tek® O.C.T compound), thereafter quickly frozen in isopentane cooled by liquid nitrogen. These specimens were subsequently stored at -80°C until sectioning commenced.
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